Intermediates for inhibitors of HIV protease and method of preparation thereof

ABSTRACT

The present invention provides novel HIV protease inhibitors, pharmaceutical formulations containing those compounds and methods of their use.

CROSS REFERENCE TO RELATED APPLICATION

This application is a division of application Ser. No. 07/875,908, filedApr. 29, 1992, now U.S. Pat. No. 5,508,409 which is acontinuation-in-part of application Ser. No. 07/727,787, filed Jul. 10,1991 (now abandoned).

BACKGROUND OF THE INVENTION

A retrovirus designated human immuno-deficiency virus (HIV) is thecausative agent of the complex disease termed Acquired Immune DeficiencySyndrome (AIDS), and is a member of the lentivirus family ofretroviruses (M. A. Gonda, F. Wong-Staal NR. C. Galo, "Sequence Homologyand Morphological Similarity of HTLV III And Visna Virus, A PathogenicLentivirus", Science, 227, 173, (1985); and P. Sonigo and N. Alizon, etal., "Nucleotide Sequence of the Visna Lentivirus: Relationship to theAIDS Virus", Cell, 42,369, (1985)). The complex disease AIDS includesprogressive destruction of the immune system and degeneration of thecentral and peripheral nervous systems. The HIV virus was previouslyknown or referred to as LAV, HTLV-III, or ARV.

A common feature of retrovirus replication is the post-translationalprocessing of precursor polyproteins by a virally encoded protease togenerate mature viral proteins required for viral assembly and function.Interruption of this processing appears to prevent the production ofnormally infectious virus. Unprocessed structural proteins also havebeen observed in clones of non-infectious HIV strains isolated fromhuman patients. The results suggest that inhibition of the HIV proteaserepresents a viable method for the treatment of AIDS and the preventionor treatment of infection by HIV.

The HIV genome encodes structural protein precursors known as gag andpol, which are processed to afford the protease, reverse transcriptaseand endonuclease/integrase. The protease further cleaves gag and gag-polpolyproteins to yield mature structural proteins of the virus core.

Considerable efforts are being directed toward the control of HIV bymeans of the structural protein precursors which are processed to yieldthe retroviral protease, reverse transcriptase andendonuclease/integrase. For example, the currently used therapeutic,AZT, is an inhibitor of the viral reverse transcriptase (H. Mitsuya, NS.Broder, "Inhibition of the In Vitro Infectivity in Cytopathic Effects ofHTLV III", Proc. Natl. Acad. Sci. USA, 83, 1911 (1986).

Research efforts have also been directed toward HIV protease inhibitors.For example, EPA 361 341; EPA 346 847; EPA 402 646; and EPA 337 714 alldisclose compounds which are said to be useful as HIV proteaseinhibitors.

Unfortunately, many of the known compounds suffer from toxicityproblems, lack of bioavailability or are short lived in vivo.

Despite the recognized therapeutic potential associated with a proteaseinhibitor and the research efforts expended thus far, a viabletherapeutic agent has not yet emerged.

Accordingly, a primary object of the present invention is to providenovel HIV protease inhibitors which are useful in the treatment of AIDS.

A further object of the present invention is to provide therapeuticcompositions that are of value in the prevention and/or treatment ofinfection by HIV and the treatment of the resulting acquired immunedeficiency syndrome.

Still another object is to provide methods for the prevention and/ortreatment of infection by HIV and the resulting acquired immunedeficiency syndrome.

Other objects, features, and advantages will become apparent to thoseskilled in the art from the following description and claims.

SUMMARY OF THE INVENTION

The present invention relates to compounds and pharmaceuticallyacceptable salts thereof that inhibit the protease encoded by humanimmunodeficiency virus (HIV) type 1 (HIV-1) and type 2 (HIV-2). Thesecompounds are useful in the prevention of infection by HIV, thetreatment of infection by HIV and/or the treatment of the resultingacquired immune deficiency syndrome (AIDS) either as compounds,pharmaceutically acceptable salts, pharmaceutical compositioningredients, whether or not in combination with other anti-virals,immunomodulators, antibiotics or vaccines. Methods of treating AIDS,methods of preventing infection by HIV and methods of treating infectionby HIV are also disclosed.

The compounds of the present invention are those having the Formula:##STR1## where:

R is C₅ -C₇ cycloalkyl, heterocycle, aryl or unsaturated heterocycle;

X is a bond, (--CH₂ --)_(q), --O--(--CH₂ --)_(q) --, --(--CH₂ --)_(q)--O-- or --N(R⁵) (CH₂ --)_(m) --;

n is 0, 1, or:2;

q is 1, 2, 3 or 4;

R¹ is aryl or C₅ -C₇ cycloalkyl;

R² is an amino acid side chain, unsaturated heterocycle, unsaturatedheterocycle (C₁ -C₄ alkanediyl), C₁ -C₄ alkylaminocarbonyl (C₁ -C₄alkanediyl), or a group having the structure --CH₂ --C(O)--NR⁴ --X--R or--CH₂ --R;

Y is an azyl or unsaturated heterocycle;

R³ is a group having the structure: ##STR2## where: 1 is 3, 4 or 5;

m at each occurrence is independently 0, 1, 2, or 3;

p is 4 or 5;

R⁴ at each occurrence is independently hydrogen, C₁ -C₆ alkyl orhydroxy(C₁ -C₄)alkanediyl;

R⁵ and R⁶ are independently selected from hydrogen, hydroxy, C₁ -C₆alkyl, C₁ -C₆ alkoxy, amino, C₁ -C₄ alkylamino, hydroxy(C₁-C₄)alkanediyl, carboxy, (C₁ -C₄ alkoxy)carbonyl, aminocarbonyl, C₁ -C₄alkylaminocarbonyl, aryl, heterocycle or unsaturated heterocycle; or apharmaceutically acceptable salt or solvate thereof.

This invention also provides pharmaceutical formulations which comprisea compound of Formula I, or the pharmaceutically acceptable salt orsolvate thereof, in association with a pharmaceutically acceptablecarrier, diluent or excipient.

A further embodiment of the present invention is a method for inhibitingHIV protease. More particularly, the present invention contemplates amethod for treating infection by HIV comprising administering to amammal in need of HIV inhibition, an HIV inhibiting dose of a compoundof Formula I or a pharmaceutically acceptable salt or solvate thereof.

A further embodiment of the present invention is a class of novelintermediates useful for preparing compounds of Formula I and a processfor preparing said intermediates. The intermediates have the Formula##STR3## where R¹ is aryl or C₅ -C₇ cycloalkyl;

R^(b) is hydrogen or an amino protecting group;

Y is aryl or unsaturated heterocycle;

R³ is a group selected from: ##STR4## where R⁴, R⁵, R⁶, 1 and p are asdefined above for Formula I or a pharmaceutically acceptable salt orsolvate thereof.

The process aspect of the present invention is a process for preparingthe ketone intermediates of Formula II which comprises:

a) reacting a compound having the formula: ##STR5##

where R³ and Y are as defined above for Formula I with a C₁ -C₄ alkyllithium or lithium di(C₁ -C₄ alkyl) amide base either in the presence orabsence of a tetramethyl (C₁ -C₄ alkylene) diamine catalyst in anaprotic solvent to afford the corresponding anion; and

b) reacting the anion from (a) with an amide having the Formula ##STR6##

where R^(b) and R¹ are as defined above for Formula I in an aproticsolvent to afford said ketone intermediate.

DETAILED DESCRIPTION OF THE INVENTION

The term "alkyl" by itself or as part of another substituent, unlessotherwise stated, includes straight or branched chain groups such asmethyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl,sec-butyl, and where indicated, higher homologs and isomers such asn-pentyl, n-hexyl, 2-methylpentyl and the like. The term "alkoxy"represents an alkyl group of the stated number of carbon atoms attachedthrough an oxygen bridge such as methoxy, ethoxy, n-propoxy, isopropoxyand the like. The term "hydroxy(C₁ -C₄) alkanediyl" means a divalentalkyl group having the stated number of carbon atoms bonded to a hydroxygroup, such as hydroxymethyl, hydroxyethyl, hydroxypropyl, andhydroxybutyl. The term "C₁ -C₄ alkylamino" means a group (--NH(C₁ -C₄alkyl)) where the alkyl group has the stated number of carbon atoms. Theterm "C₁ -C₄ dialkylamino" means a group (--N(C₁ -C₄ alkyl)₂) where eachalkyl group, independently, has the stated number of carbon atoms. Theterm "cycloalkyl" means a saturated ring group having the stated numberof carbon atoms such as cyclopentyl, cyclohexyl and cycloheptyl.

The term "heterocycle" means an unsubstituted or substituted stable 5-to 7-membered monocyclic and stable 7- to 10-membered bicyclicheterocyclic ring which is saturated and which consists of carbon atomsand from one to three heteroatoms selected from the group consisting ofN, O, S, and wherein the nitrogen and sulfur heteroatoms may optionallybe oxidized, and the nitrogen heteroatom may optionally be quaternizedand including a bicyclic group in which any of the above-definedheterocyclic rings is fused to a benzene ring. The heterocyclic ring maybe attached at any heteroatom or carbon atom which affords a stablestructure. The heterocycle is unsubstituted or substituted with 1, 2, or3 substituents independently selected from halo, C₁ -C₅ alkyl, C₁ -C₅alkoxy, carboxy, (C₁ -C₄ alkoxy)carbonyl, aminocarbonyl, C₁ -C₄alkylaminocarbonyl, amino, C₁ -C₄ alkylamino, C₁ -C₄ dialkylamino or agroup --(CH₂)_(q) --R⁷ where q is 1, 2, 3, or 4 and R⁷ is hydroxy, C₁-C₄ alkoxy, carboxy, C₁ -C₄ alkoxycarbonyl, amino, aminocarbonyl, C₁ -C₄alkylamino or C₁ -C₄ dialkylamino.

The term "unsaturated heterocycle" means an unsubstituted or substitutedstable 5- to 7-membered monocyclic and stable 7- to 10-membered bicyclicheterocyclic ring which has one or more double bonds and which consistsof carbon atoms and from one to three heteroatoms selected from thegroup consisting of N, o, S, and wherein the nitrogen and sulfurheteroatoms may optionally be oxidized, and the nitrogen heteroatom mayoptionally be quarternized and including a bicyclic group in which anyof the above-defined heterocyclic rings is fused to a benzene ring. Theunsaturated heterocyclic ring may be attached at any heteroatom orcarbon atom which affords a stable structure. The unsaturatedheterocycle is unsubstituted or substituted with 1, 2, or 3 substituentsindependently selected from halo, C₁ -C₅ alkyl, C₁ -C₅ alkoxy, carboxy,(C₁ -C₄ alkoxy)carbonyl, aminocarbonyl, C₁ -C₄ alkylaminocarbonyl,amino, C₁ -C₄ alkylamino, C₁ -C₄ dialkylamino or a group --(CH₂)_(q)--R⁷ where q is 1, 2, 3, or 4 and R⁷ is hydroxy, C₁ -C₄ alkoxy, carboxy,C₁ -C₄ alkoxycarbonyl, amino, aminocarbonyl, C₁ -C₄ alkylamino or C₁ -C₄dialkylamino.

Examples of such heterocycles and unsaturated heterocycles includepiperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl,2-oxo-pyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl,pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl,imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl,oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl,thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazolyl, benzopyranyl,benzothiazolyl, benzoazolyl, furyl, tetrahydrofuryl, tetrahydropyranyl,thienyl, benzothienyl, thiamorpholinyl, thiamorpholinylsulfoxide,thiamorpholinylsulfone, oxadiazolyl, triazolyl, tetrahydroquinolinyl andtetrahydrisoquinolinyl.

The term "aryl" means unsubstituted or substituted phenyl orunsubstituted or substituted naphthyl. The phenyl or naphthyl ring isunsubstituted or substituted with 1, 2, or 3 substituents independentlyselected from halo, C₁ -C₅ alkyl, C₁ -C₅ alkoxy, carboxy, (C₁ -C₄alkoxy)carbonyl, aminocarbonyl, C₁ -C₄ alkylaminocarbonyl, amino, C₁ -C₄alkylamino, C₁ -C₄ dialkylamino, benzyloxy or a group --(CH₂)_(q) --R⁷where q is 1, 2, 3, or 4 and R⁷ is hydroxy, C₁ -C₄ alkoxy, carboxy, C₁-C₄ alkoxycarbonyl, amino, aminocarbonyl, C₁ -C₄ alkylamino or C₁ -C₄dialkylamino. The term "halogen" and "halo" mean any of chloro, bromo,fluoro and iodo.

The third group in the definition of R³ includes unsubstituted andsubstituted piperidinyl, and unsubstituted and substituted pyrrolidinylwhere the substituents are selected from those defined for R⁵ and R⁶which affords a sterically feasible stable structure.

The term "amino acid side chains" means the distinctive atom or groupbonded to an α-carbon atom also having bonded thereto a carboxyl groupand an amino group. These side chains are selected from those found on Dand L:

    ______________________________________                                               Alanine            Ala                                                        Arginine           Arg                                                        Asparagine         Asn                                                        Aspartic acid      Asp                                                        Cysteine           Cys                                                        Glutamine          Gln                                                        Glutamic acid      Glu                                                        Glycine            Gly                                                        Histidine          His                                                        Isoleucine         Ile                                                        Leucine            Leu                                                        Lysine             Lys                                                        Methionine         Met                                                        Phenylalanine      Phe                                                        Proline            Pro                                                        Serine             Ser                                                        Threonine          Thr                                                        Tryptophan         Trp                                                        Tyrosine           Tyr                                                        Valine             Val                                                 ______________________________________                                    

The term "amino-protecting group" as used in the specification refers tosubstituents of the amino group commonly employed to block or protectthe amino functionality while reacting other functional groups on thecompound. Examples of such amino-protecting groups include the formylgroup, the trityl group, the phthalimido group, the trichloroacetylgroup, the chloroacetyl, bromoacetyl and iodoacetyl groups,urethane-type blocking groups such as benzyloxycarbonyl,4-phenylbenzyloxycarbonyl, 2-methylbenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl,4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl,2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyloxycarbonyl,4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl,4-nitrobenzyloxycarbonyl, 4-cyanobenzyloxy-carbonyl,2-(4-xenyl)isopropoxycarbonyl, 1,1-diphenyleth-1-yloxycarbonyl,1,1-diphenylprop-1-yloxycarbonyl, 2-phenylprop-2-yloxycarbonyl,2-(p-toluyl)prop-2-yloxycarbonyl, cyclopentanyloxycarbonyl,1-methylcyclopentanyloxycarbonyl, cyclohexanyloxycarbonyl,1-methylcyclohexanyloxycarbonyl, 2-methylcyclohexanyloxycarbonyl,2-(4-toluylsulfonyl)ethoxycarbonyl, 2(methylsulfonyl)ethoxycarbonyl,2-(triphenylphosphino)ethoxycarbonyl, fluorenylmethoxycarbonyl ("FMOC"),2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl,1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl,5-benzisoxalylmethoxycarbonyl, 4-acetoxybenzyloxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl,cyclopropylmethoxycarbonyl, 4-(decyloxy)benzyloxycarbonyl,isobronyloxycarbonyl, 1-piperidyloxycarbonyl and the like; thebenzoylmethylsulfonyl group, the 2-(nitro)phenylsulfenyl group, thediphenylphosphine oxide group and like amino-protecting groups. Thespecies of amino-protecting group employed is not critical so long asthe derivatized amino group is stable to the condition of subsequentreaction(s) on other positions of the intermediate molecule and can beselectively removed at the appropriate point without disrupting theremainder of the molecule including any other amino-protecting group(s).Preferred amino-protecting groups are t-butoxycarbonyl, andbenzyloxycarbonyl. Further examples of groups referred to by the aboveterms are described by J. W. Barton, "Protective Groups in OrganicChemistry", J. G. W. McOmie, Ed., Plenum Press, New York, N.Y., 1973,Chapter 2, and T. W. Greene, "Protective Groups in Organic Synthesis",John Wiley and sons, New York, N.Y., 1981, Chapter 7.

The compounds of the present invention have at least three asymmetriccenters denoted by an asterisk in Formula IA below. ##STR7##

As a consequence of these chiral centers, the compounds of the presentinvention occur as racemates, racemic mixtures and as individualdiastereomers. All asymmetric forms, individual isomers and combinationsthereof, are within the scope of the present invention.

As mentioned above, the invention includes pharmaceutically acceptablesalts of the compounds defined by the above formula. Although generallyneutral, a particular compound of this invention can possess asufficiently acidic, a sufficiently basic, or both functional groups,and accordingly react with any of a number of nontoxic inorganic bases,and nontoxic inorganic and organic acids, to form a pharmaceuticallyacceptable salt. Acids commonly employed to form acid addition salts areinorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodicacid, sulfuric acid, phosphoric acid, and the like, and organic acidssuch as p-toluene-sulfonic, methanesulfonic acid, oxalic acid,p-bromo-phenylsulfonic acid, carbonic acid, succinic acid, citric acid,benzoic acid, acetic acid, and the like. Examples of suchpharmaceutically acceptable salts thus are the sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide,iodide, acetate, propionate, decanoate, caprylate, acrylate, formate,isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate,succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate,hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate,dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate,xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate,citrate, lactate, g-hydroxybutyrate, glycollate, tartrate,methanesulfonate, propanesulfonate, naphthalene-1-sulfonate,napththalene-2-sulfonate, mandelate and the like. Preferredpharmaceutically acceptable acid addition salts are those formed withmineral acids such as hydrochloric acid and hydrobromic acid, and thoseformed with organic acids such as maleic acid and methanesulfonic acid.

Base addition salts include those derived from inorganic bases, such asammonium or alkali or alkaline earth metal hydroxides, carbonates,bicarbonates, and the like. Such bases useful in preparing the salts ofthis invention thus include sodium hydroxide, potassium hydroxide,ammonium hydroxide, potassium carbonate, sodium carbonate, sodiumbicarbonate, potassium bicarbonate, calcium hydroxide, calciumcarbonate, and the like. The potassium and sodium salt forms areparticularly preferred.

Preferred compounds of this invention are those of Formula I where

R is aryl or unsaturated heterocycle;

X is a bond, --O--(--CH₂ --)_(q) --, or --(--CH₂ --)_(q) --O--;

q is 1 or 2;

n is 1 or 2;

R¹ is aryl;

R² is an amino acid side chain or unsaturated heterocycle (C₁ -C₄alkanediyl);

Y is aryl;

R³ is --C(O)--NR₄ R⁴ or --N(R₅)C(O)--R⁶

where R⁴, R⁵ and R⁶ are independently and at each occurrence hydrogen orC₁ -C₆ alkyl; or a pharmaceutically acceptable salt or solvate thereof.

Alternatively, preferred compounds of this invention are those ofFormula I where:

R is aryl or unsaturated heterocycle;

X is bond, (--CH₂ --)_(q), --O--(--CH₂ --)_(q) --, or --(--CH₂ --)_(q)--O--;

q is 1 or 2;

n is 1 or 2;

R¹ is aryl;

R² is an amino acid side chain, unsaturated heterocycle (C₁ -C₄alkanediyl) or --CH₂ --C(O)--NR⁴ --X--R;

Y is aryl;

R³ is --C(O)--NR₄ R⁴ or --N(R₅)C(O)--R⁶ where R⁴, R⁵ and R⁶ areindependently and at each occurrence hydrogen or C₁ -C₆ alkyl; or apharmaceutically acceptable salt or solvate thereof.

More preferred compounds are those of Formula I where:

R is naphthyl, quinoxalinyl or quinolinyl, each of said radicalsunsubstituted or substituted with one or two C₁ -C₄ alkyl groups;

X is a bond, --OCH₂ ----CH₂ O--;

n is 1;

R¹ is phenyl;

R² is --CH₂ --C(O)--NH₂ ;

Y is phenyl, unsubstituted or substituted with C₁ -C₄ alkyl; or thepharmaceutically acceptable salt or solvate thereof.

Particularly preferred compounds are those of Formula IB: ##STR8## whereat each occurence R⁴ is independently hydrogen or C₁ -C₄ alkyl; or apharmaceutically acceptable salt or solvate thereof.

The compounds of the present invention, or their precursors, areprepared using procedures known to those of ordinary skill in art. Moreparticularly, the compounds of Formula I where R³ is bonded through acarbonyl group (groups 1 through 3 in the definition of R³) are preparedaccording to the procedures shown below in Scheme 1 and as describedfollowing Scheme 1. ##STR9## where:

R, X, n, R¹, R², and Y are as defined above for Formula I, R^(b) is anamino protecting group, and R^(3a) is group 1 through 3 of R³ as definedabove for Formula I.

In Scheme 1, a suitable aryl or unsaturated heterocycle carboxylic acidis activated, that is, converted, in Reaction 1 to the correspondingacid chloride or acid bromide by reaction with thionyl chloride, thionylbromide, phosphorous trichloride, phosphorous tribromide, phosphorouspentabromide or phosphorous pentachloride according to procedures andunder conditions well known to those skilled in the art. The acidchloride or acid bromide afforded by Reaction 1 can be isolated orfurther reacted as shown in Reaction 2. Suitable aryl or unsaturatedheterocycle carboxylic acid compounds are commercially available orprepared by standard procedures well known to those skilled in the art.

In Reaction 2, the acid chloride or acid bromide is reacted with aprimary or secondary amine having the formula R³ --H where R³ is asdefined above for Formula I in a nonpolar aprotic solvent or mixture ofsolvents in the presence or absence of an acid scavenger at atemperature of from about -20° C. to about 25° C. to afford thecorresponding amide. Suitable solvents for this reaction include ethersand chlorinated hydrocarbons, preferably diethyl ether, chloroform, ormethylene chloride. Preferably, this reaction is carried out in thepresence of an acid scavenger such as a tertiary amine, preferablytriethyl amine. The amide afforded by this reaction may be isolated orfurther reacted as shown in Reaction 3.

In Reaction 3, the amide is reacted with a strong base in the presenceof a catalyst in an aprotic solvent at a temperature of from about -78°C. to about 0° C. to afford the corresponding anion which is reactedwith a Weinreb Amide in an aprotic solvent at a temperature from about-80° C. to about -40° C. to afford a ketone which may be isolated orfurther reacted as shown in Reaction 5. Suitable bases for Reaction 3include lithium amide bases and alkyl lithium bases, preferably C₁ -C₄alkyl lithium bases and lithium di(C₁ -C₄ alkyl)amide bases. Suitablesolvents for Reaction 3 are ethers and preferably tetrahydrofuran (THF).Suitable catalysts for Reaction 3 are tetramethyl (C₁ -C₄ alkylene)diamines and preferably tetramethylethylenediamine. In Reaction 4 about2 equivalents of the anion are used per equivalent of Weinreb Amide.

In Reaction 5 the ketone is reduced using a suitable reducing agent in aprotic solvent at a temperature of from about -25° C. to about 25° C. tothe corresponding alcohol. Suitable reducing agents for this reactioninclude sodium borohydride, lithium borohydride, diisobutylaluminumhydride, and sodium bis(2-methoxyethoxy)aluminum hydride, preferablysodium borohydride. Suitable protic solvents for this reaction includealcohols, and preferably ethanol.

Reaction 6 is a standard amino deprotection reaction using proceduresand methods well known to those skilled in the art to afford thecorresponding amine which may be isolated or further reacted in Reaction7.

Reaction 7 is a standard coupling reaction commonly employed in thesynthesis of peptides where an amine is reacted with a carboxylic acidin an aprotic solvent or mixture of solvents in the presence or absenceof a catalyst, preferably in the presence of a catalyst, and in thepresence of a coupling reagent. Suitable aprotic solvents for thisreaction are tetrahydrofuran and dimethylformamide, and preferably amixture of such solvents, at a temperature from about -30° C. to about25° C. A catalyst is preferably included for this reaction and thepreferred catalyst is hydroxybenzotriazole. Examples of suitablecoupling reagents include the carbodiimides such as N,N¹-diethylcarbodiimide; the imidazoles such as carbonyldiimidazole; aswell as reagents such as 1-hydroxybenzotriazole mesylate orN-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). The couplingreaction is carried out by adding about an equimolar quantity of theamine to a solution of the carboxylic acid in the presence of anequimolar quantity to slight excess of the coupling reagent. Thepreferred coupling reagent for this reaction is dicyclohexylcarbodiimide. The coupling reaction affords the compounds of Formula Ior their precursors.

Alternatively, the divalent .paren open-st.NH--CHR² --C(O).parenclose-st._(n) moiety can be coupled with the intermediate afforded byReaction 6 and then coupled to an R--X--COOH to afford compounds ofFormula I, where n, R, X and R₂ are as defined above for Formula I. Insuch a case, the reactant Cb_(z) .paren open-st.NH--CHR² --C(O).parenclose-st._(n) OH is reacted with the intermediate afforded by Reaction 6under standard coupling reaction conditions well known to those skilledin the art and commonly employed in the synthesis of peptides. The aminoprotecting group, CB_(z) is shown and preferred, but others may also beemployed, is removed again using standard amino deprotection reactionconditions well known to those skilled in the art to afford thecorresonding intermediate having a terminal primary amino reactivegroup. A further coupling reaction is carried out with the R--X--COOHreactant again under standard coupling reaction conditions well known tothose skilled in the art and commonly employed in the synthesis ofpeptides, to afford the compounds of Formula I.

In addition, for those compounds where X is --N(R⁵)(CH₂ --)_(m) --, theamine intermediate may be reacted with compounds of the formula, R--(CH₂--)_(m) --N═C═O to afford compounds of Formula I, where R, m and R⁵ areas defined above for Formula I. The isocyanate reactant, R--(CH₂ --)_(m)--N═C═O, to the extent not commercially available, is prepared bystandard methodology well known to those skilled in the art. Thereaction is carried out by combining an equimolar quantity of the amineintermediate with an equimolar quantity to a slight excess of theisocyanate reactant. The reaction is carried out in an aprotic solvent,at a temperature from about 15° C. to about 35 C, and preferably underan inert atmosphere, such as nitrogen. Suitable aprotic solvents forthis reaction are tetrahydrofuran and acetonitrile. The reaction affordscompounds of Formula I, wherein X is --N(R⁵)(CH₂ --)_(m) --.

The Weinreb Amide used as a reactant in Reaction 4 is prepared byreacting an amino-protected amino acid with N-methoxy-N-methyl-amine inthe presence of a catalyst, an acid scavenger, and a coupling agent andpreferably in the presence of an emulsifier in an aprotic solvent ormixture of solvents at a temperature of from about -25° C. to 25° C. Thepreferred catalyst for this reaction is hydroxybenzotriazole. Thepreferred acid scavenger is a tertiary alkylamine and preferablytriethylamine. The preferred emulsifier for this reaction isN-methylmorpholine. The preferred coupling reagent is ethyldimethylaminopropylcarbodiimide hydrochloride. The Weinreb Amideafforded by this reaction is preferably isolated prior to its use inReaction 4 of Scheme 1.

The acid reactant in Reaction 7, to the extent not commerciallyavailable, is prepared by standard methodology well known to thoseskilled in the art.

Suitable amino-protecting groups are those substituents of an aminogroup commonly employed by those skilled in the art to block or toprotect the amino functionality while reacting other functional groupson the compound as defined above. Preferred amino-protecting groups aret-butoxycarbonyl (Boc) and benzyloxycarbonyl (cbz).

The compounds of Formula I where R³ is bonded through a nitrogen atom(groups 4 through 6 in the definition of R³) are prepared according tothe procedures shown below in Scheme 2 and as described following Scheme2. ##STR10## wherein R, X, n, R¹, R² and Y are as defined above forFormula I, R^(b) is an amino protecting group, and R^(3b) is group 4through 6 of R³ as defined above for Formula I.

In Scheme 2, a suitable aryl or unsaturated heterocycle amine isamino-protected, as necessary, in Reaction 1 under standard conditionswell known to those skilled in the art. Reactions 2 through 6 arecarried out substantially as described above in Scheme 1 for Reactionsthrough 7. In Scheme 2, a further deprotective reaction, Reaction 7, isnecessary to remove the amino protecting group introduced in Reaction 1.This is a standard amino deprotection reaction using procedures andmethods well known to those skilled in the art. For example, the Bocgroup illustrated in Reaction 1, Scheme 2, is removed using a strongacid, preferably trifluoroacetic acid. Finally, in Reaction 8, theillustrated intermediate is acylated with a suitable acid chloride,isocyanate or chloroformate, preferably in the presence of an acidscavenger such as triethylamine.

Alternatively, the divalent moiety .paren open-st.N--H--CHR₂--C(O)--.paren close-st._(n) portion of the reactant illustrated forReaction 6 can be coupled to the intermediate afforded by Reaction 5substantially as described above for Scheme 1, where n and R² are asdefined above for Formula I. The coupling reaction with an R--X--COOHreactant, where R and X areas defined above for Formula I, can becarried out prior to or subsequent to the acylation step, Reaction 8. Ineither instance, deprotection of the primary amine functional group onthe corresponding intermediate is required prior to the coupling oracylation reactions. To facilitate deprotection of the desired primaryamine, the amino protecting groups employed should not both be removableunder substantially similar conditions. Such deprotection is carried outby standard techniques and under standard conditions for such reactionswell known to those skilled in the art. As described above for Scheme 1,the coupling reaction with an R--X--COOH is, again, a standard couplingreaction carried out under conditions well known to those skilled in theart and commonly employed in the synthesis of peptides.

As noted above, the optically active diastereomers of the compounds ofFormula I are considered part of this invention. Such optically activeisomers may be prepared from their respective optically activeprecursors by the procedures described above, or by resolving theracemic mixtures. The resolution can be carried out in the presence of aresolving agent, by chromatography or by repeated crystallization or bysome combination of these techniques which are known to those skilled inthe art. Further details regarding resolutions can be found in Jacques,et al., Enantiomers, Racemates, and Resolutions, john Wiley & Sons 1981.

The compounds employed as initial starting material in the synthesis ofthe compounds of this invention are well known and, to the extent notcommercially available are readily synthesized by standard procedurescommonly employed by those of ordinary skill in the art.

The pharmaceutically acceptable salts of the invention are typicallyformed by reacting a compound of Formula I with an equimolar or excessamount of acid or base. The reactants are generally combined in a mutualsolvent such as diethyl ether or benzene, for acid addition salts, orwater or alcohols for base addition salts, and the salts normallyprecipitate out of solution within about one hour to about ten days andcan be isolated by filtration or other conventional methods.

In addition some of the compounds of Formula I may form solvates withwater or with common organic solvents. Such solvates are included withinthe scope of the compounds of the present invention.

The following Preparations and Examples further illustrate the compoundsof the present invention and methods for the synthesis. The examples arenot intended to be limiting to the scope of the invention in any respectand should not be so construed.

Unless otherwise noted, NMR data appearing in the examples refers to thefree base of the subject compound.

In the following Preparations and Examples, the terms melting point,nuclear magnetic resonance spectra, electron impact mass spectra, fielddesorption mass spectra, East atom bombardment mass spectra, infraredspectra, ultraviolet spectra, elemental analysis, specific rotation,high performance liquid chromatography, and thin layer chromatographyare abbreviated m.p., n.m.r., m.s., f.d.m.s., f.a.b.m.s., i.r., u.v.,anal., o.r., HPLC, and TLC, respectively. In addition, the absorptionmaxima listed for the i.r. spectra are only those of interest and notall of the maxima observed.

In conjunction with n.m.r. spectra, the following abbreviations areused: "s" is singlet, "d" is doublet, "dd" is doublet of doublets, "t"is triplet, "q" is quartet, "m" is multiplet, "dm" is a doublet ofmultiplets and "br.s", "br.d", "br.t", and "br.m" are broad singlet,doublet, triplet, and multipier respectively. "J" indicates the couplingconstant in Hertz. "DMSO-d₆ " is dimethyl sulfoxide where protons havebeen replaced with deuterium.

The n.m.r. spectra were obtained on a Varian Associates EM-390 90 MHz orT-60 60 MHz instrument, on a Jeol FX-90Q 90MHz instrument, on a BrukerCorp. 270 MHz instrument or on a General Electric QE-300 300 MHzinstrument. The chemical shifts are expressed in delta values (parts permillion downfield from tetramethylsilane). The field desorption massspectra were taken on a Varion-MAT 731 Spectrometer using carbondendrite emitters. Electron Impact Mass Spectra were obtained on a CEC21-110 instrument from Consolidated Electrodynamics Corporation.Infrared spectra were obtained on a Perkin-Elmer 281 instrument.Ultraviolet Spectra were obtained on a Cary 118 instrument. Specificrotations were obtained on a Perkin-Elmer Q-41 instrument. Thin layerchromatography was carried out on E. Merck silica gel plates. Meltingpoints are uncorrected.

PREPARATION 1 Preparation of(S)-N-methoxy-N-methyl-2-(N-phenylmethyloxycarbonyl)amino-3-phenylpropanamide

A. 1 L, single necked round bottom flask was charged with 50.35 g (0.17mol) cbz-L-phenylalanine in 600 mL methylene chloride (CH₂ Cl₂). To thissolution was added 21.3 g (0.22 mol) N,O-dimethylhydroxylamine, 30 mL(22 g, 0.22 mol) triethylamine (TEA), 29.6 g (0.22 mol)1-hydroxybenztriazole hydrate (HOBT), and 37 mL (34 g, 0.34 mol)N-methylmorpholine. The resulting solution was cooled to 0° C. and 35.5g (0.19 mol) 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(EDC) was added in one portion. The reaction mixture was stirred at 0°C. for one hour, and then warmed to room temperature and stirredovernight. The solution was then diluted with 900 mL hexane andtransferred to a separatory funnel, where it was washed twice with 500mL saturated NaHCO₃, twice with 500 mL 1M NaHSO₄, once with 500 mLbrine, and dried over Na₂ SO₄. Removal of the drying agent by filtrationand concentration in vacuo afforded a viscous colorless oil (56.89 g,97%).

a!_(D) +11.33° (c 1.059, MeOH).

IR (CHCl₃) 3435, 3027, 3013, 1717, 1659, 1507, 1455, 1392, 1231, 1051cm⁻¹.

MS (FD) m/e 342(M+), 342(100).

Analytical calc'd for C₁₉ H₂₂ N₂ O₄ : C 66.65, H 6.48, N 8.18; found C66.58, H 6.59, N 8.20.

PREPARATION 2 Preparation of quinaldic acid pentafluorophenyl ester

A. An oven dried single necked round bottom flask was charged with 15.0g quinaldic acid (86.6 mmol), 20.8 g pentafluorophenol (113 mmol), and200 mL THF. The suspension was stirred, and 18.3 g EDC (95.3 mmol) wasadded in one portion. Vigorous stirring was continued at roomtemperature for two hours, during which time a gummy precipitate formedat the bottom of the flask. The solution was decanted from the gum, andthe gum was washed with CH₂ C₁₂. The combined organic layers werediluted with hexane and washed once with 50 mL 0.1N NaHSO₄, twice with50 mL 1N K₂ CO₃, and once with 50 mL brine. The organic layer was driedover Na₂ SO₄, filtered, and concentrated in vacuo, affording a pale pinksolid. The solid was dissolved in 30 mL hot diethyl ether and 400 mL hothexane was added. The solution was allowed to cool to room temperature,and then to 0° C. over a period af 1.5 hours. The desired product wascollected as colorless needles (21.6 g, 73%) by filtration.

¹ H NMR (300 MHz, CDCl₃) d 7.73 (t, J=7.5 Hz, 1H), 7.86 (t, J=7.9 Hz,1H), 7.95 (d, J=8.2 Hz, 1H), 8.29-8.42 (m, 3H).

IR (CHC₁₃) 3035, 2997, 1763, 1522, 1285, 1068, 998, 842 cm⁻¹.

Analytical calc'd for C₁₆ H₆ NO₂ F₅ : C 56.65, H 1.78, N 4.13; found C56.66, H 1.77, N 4.12.

PREPARATION 3 Preparation of(S)-2-(2-N-quinolinylcarboxy)-2,4-diamino-1,4-butanedioic acid

B. A 1L round bottom flask was charged with 17.9 g quinaldic acidpentafluorophenyl ester (51.2 mmol), 6.99 g L-asparagine monohydrate(46.6 mmol), 15.7 g sodium bicarbonate (186 mmol), 265 mL water and 219mL dioxane. The resulting suspension was stirred vigorously overnight atroom temperature, during which time >90% of the reaction mixture wassolubilized. The reaction mixture was concentrated in vacuo to removethe dioxane, and the resulting aqueous layer acidified to pH 3 with 2NNaHSO₄. The aqueous layer was then extracted three times with 60 mL 3:1chloroform (CHCl₃)/isopropanol (i-PrOH). The combined organic layerswere washed once with 50 mL brine and dried over Na₂ SO₄. Removal of thedrying agent by filtration followed by concentration under reducedpressure provided a colorless solid, which was washed with 500 mLdiethyl ether and 250 mL hot hexanes to remove residualpentafluorophenol. The remaining solid was dried at 80° C. in a vacuumoven for three hours, giving 10.61 g (79%) of the desired product.

a!_(D) +16.54° (c 1.01, DMSO).

¹ H NMR (300 MHz, DMSO-D₆) d 2.68 (dd, J=16.0, 4.9 Hz, 1H), 2.81 (dd,J=16.0, 5.7 Hz, 1H), 4.74-4.81 (m, 1H), 6.96 (s, 1H), 7.70 (t, J=7.5 Hz,1H), 7.85 (t, J=7.5 Hz, 1H), 8.05-8.15 (m, 3H), 8.56 (d, J=8.5 Hz, 1H),9.12 (d, J=8.6 Hz, 1H), 12.8 (s, 1H).

IR (KBr) 3385, 3367, 3216, 1171, 1662, 1523, 1499, 1427, 780, 592 cm⁻¹.

MS (FD) m/e 288 (M+) , 288 (100).

Analytical calc'd for C₁₄ H₁₃ N₃ O₄ : C 58.53, H 4.56, N 14.63; found C58.80, H 4.57, N 14.56.

EXAMPLE 1 1S-(1R*, 4R*.5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide ##STR11## Preparation of N-t-butyl-2-methylbenzamide

A three-necked 3-L round bottom flask fitted with a thermometer,mechanical stirrer and nitrogen inlet was charged with 139.2 g (0.9moles, 1.0 equiv.) of o-toluoyl chloride in 1200 mL of methylenechloride (CH₂ Cl₂) at 25° C. under a flow af nitrogen. Upon dissolving,the stirring solution, under a static atmosphere, was cooled to 0° C.with an ice/salt bath and 180.0 g of triethylamine (1.8 moles, 2.0equiv.), was added dropwise via an addition funnel over a period of 30minutes. Immediately after adding the triethylamine, a solution of 73.14g (1.0 mol, 1.1 equiv.) of t-butylamine in methylene chloride (200 mL)was added dropwise over 1.5 h. Upon completion of the addition, thereaction was allowed to warm to room temperature and stir 2.5 h. Next300 mL of water was added to the reaction mixture, and it was pouredinto a separatory funnel containing 1500 mL of water. The organic layerwas washed with 2×150 mL 2N NaOH, 1×150 mL of 1.0/HCl, and 2×250 mL ofbrine. The organic layer was dried over magnesium sulfate, filtered, andevaporated under reduced pressure to give 167.6 g (97%) of an off-whitesolid, mp 77°-78° C.

¹ H NMR (300 MHz, CDCl₃) d 1.41 (s, 9H, --C(CH₃)3), 2.41 (s, 3H, ARCH₃),5.54 (br s, 1H, --NE), 7.13-7.30 (m, 4H, aromatics).

IR (CHCl₃) 3430, 3011, 2971, 2932, 1661, 1510, 1484, 1452, 1393, 1366,1304, 1216, 876 cm ⁻¹.

MS (FD) m/e 191(M+), 191(100).

Analytical calc'd for C₁₂ H₁₇ NO: C 75.35, H 8.76, N 7.32; found C75.10, H 9.11, N 7.20.

Preparation of(S)-N-C-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-oxopropyl)benzamide

A solution of 7.0 g (36.5 mmol) N-t-butyl-2-methylbenzamide in 200 mLanhydrous tetrahydrofuran (THF) was prepared under a nitrogen atmospherein an oven dried, 500 mL pear shaped flask fitted with a rubber septum.To this solution, 12.1 mL (9.3 g, 80.3 mmol, 2.2 equiv.)N,N,N',N'-tetramethylethylenediamine (TMEDA) was added via syringe. Thesolution was cooled to -78° C., and 55.9 mL sec-butyllithium was addeddropwise through a syringe at a rate such that the internal temperatureof the reaction did not go above -60° C. The dianion solution wasallowed to stir for 1 hr. at -78° C., and then a solution of 5.00 g(14.6 mmol)(S)-N-methoxy-N-methyl-2-(N-phenylmethyl-oxycarbonyl)amino-3-phenylpropanamidein 50 mL anhydrous THF (prepared in a separate oven dried 100 mL roundbottom flask under nitrogen atmosphere) was added via cannula whilekeeping the reaction temperature below -65° C. When the addition wascomplete, the reaction mixture was allowed to warm to -20° C. andquenched with 20 mL saturated NH₄ Cl. The reaction mixture was thendiluted with 200 mL diethyl ether (Et₂ O) and transferred to aseparatory funnel, where the layers were separated. The organic layerwas washed twice with 200 mL H₂ O, once with 200 mL 0.2N NaHSO₄, oncewith 150 mL brine, and then dried over Na₂ SO₄. Removal of the dryingagent by filtration followed by concentration in vacuo gave a colorlessoil which was purified by flash chromatography with 3:1 CH₂ Cl₂ /ethylacetate (EtOAc). The product was isolated as a colorless foam (6.08 g,88%).

a!_(D) -289.26° (c 0.12, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.38 (s, 9H, --C(CH₃)₃), 2.99 (dd, J=15, 6Hz, 1H), 3.24 (dd, J=15, 6 Hz, 1H), 3.89 (d, J=18 Hz, 1H), 4.16 (d, J=18Hz, 1H), 4.72 (dd, J=15, 6 Hz, 1H), 5.00-5.09 (m, 2H), 5.56 (d, J=6 Hz,1H), 5.93 (br s, 1H), 7.03-7.40 (m, 14H, aromatics).

IR (CHCl₃) 3431, 3027, 3012, 2973, 1713, 1658, 1511, 1454, 1383, 1366,1307, 1231, 1046 cm⁻¹.

MS (FD) m/e 472(M⁺), 218(100).

Analytical calc'd for C₂₉ H₃₂ N₂ O₄ : C 73.70, H 6.82, N 5.93; found C73.41, H 6.98, N 5.83.

Preparation of R-(R*,S*)!-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-hydroxypropyl)benzamide

A solution of 6.96 g (14.7 mmol )(S)-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-oxopropyl)benzamidein 200 mL absolute ethanol was prepared in a 500 mL round bottom flaskunder a nitrogen atmosphere. To this solution was added in one portion2.78 g NaBH₄ (73.5 mmol). There was an initial slight exotherm, and themixture quickly cooled back to room temperature. The solution wasallowed to stir at room temperature, and was monitored by tlc. Afterfour hours, the reaction was complete. The ethanol solution was dilutedwith 200 mL EtOAc, and quenched by dropwise addition of 20 mL saturatedNH₄ Cl. The organic layer was then washed once with 150 mL 1N HCl, oncewith 100 mL saturated NaHCO₃, and once with 100 mL brine. Drying overNa₂ SO₄ followed by filtration and concentration under reduced pressureafforded a colorless oil (6.4 g, 93%) that was determined to be a 9:1mixture of diastereomers by ¹ H NMR. The major, desired diastereomer wasisolated by flash chromatography with a gradient of 49:1 to 9:1 ethylacetate/CH₂ Cl₂ (5.12 g, 74%).

a!_(D) +10.38° (c 0.10, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.40 (s, 9H, --C(CH₃)₃), 2.79 (dd, J=12, 3Hz, 1H), 2.90-2.98 (m, 2H), 3.04 (44, J=12, 3 Hz, 1H), 3.70-3.81 (m,1H), 3.97 (m, 1H), 4.96-5.08 (m, 2H), 5.10 (d, J=9 Hz, 1H), 5.88 (d, J=6Hz, 1H), 5.93 (s, 1H), 7.13-7.42(m, 14H).

IR (CHCl₃) 3431, 3028, 3012, 2971, 1773, 1643, 1515, 1454, 1367, 1229,1028 cm⁻¹.

MS (FD) m/e 475 (M⁺), 475 (100).

Analytical calc'd for C₂₉ H₃₄ N₂ O₄ : C 73.39, H 7.22, N 5.99; found C73.12, H 7.48, N 5.62.

Preparation of R-(R*,S*)!-N-C-butyl-2-(3-amino-2-hydroxypropyl)benzamide

A solution of 41.0 g of the R-(R*,S*)!-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-hydroxypropyl)benzamide(120 mmol) in 150 mL absolute ethanol was prepared and subjected tohydrogenation over 10% Pd/C (500 mg) in a Parr shaker apparatus. Thecatalyst was removed by filtration, and the filtrate concentrated invacuo to give 31.1 g (96%) of a light yellow foam. The foam was coupledwithout further purification to(S)-2-(2-N-quinolinylcarboxy)-2,4-diamino-1,4-butanedioic acid.

a!_(D) +34.68° (c 1.0, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.46 (s, 9H), 2.71 (dd, J=13.7, 9.5 Hz, 1H),2.84 (dd, J=13.3, 2.51 Hz, 1H), 2.95-3.06 (m, 2H), 3.23-3.29 (m, 1H),3.84-3.90 (m, 1H), 6.23 (s, 1H), 7.19-7.37 (m, 12H).

IR (CHCl₃) 3440, 3382, 3007, 2970, 2934, 1643, 1516, 1454, 1367, 1213cm⁻¹.

MS (FD) m/e 341(M⁺), 341(100).

Preparation of 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

A solution of 500 mg R-(R*,S*)!-N-C-butyl-2-(3-amino-2-hydroxypropyl)benzamide (1.46 mmol) in 10 mLanhydrous THF was prepared in a single necked 50 mL round bottom flask.To this was added 443 mg(S)-2-(2-N-quinolinylcarboxy)-2,4-diamino-1,4-butanedioic acid (1.54mmol), 208 mg HOBT•H₂ O (1.54 mmol), and 1.75 mL anhydrous DMF (enoughto obtain a homogeneous solution). The solution was cooled to -10° C. inan ice/acetone bath, and 309 mg dicyclohexylcarbodiimide (DCC) (1.50mmol) was added in one portion. The mixture was stirred under N₂ at -10°C. for 20 minutes, and then at 0° C. for 1 hour. The stirring solutionwas then allowed to warm to room temperature and stir overnight, duringwhich time the dicyclohexylurea precipitate began to form. Afterstirring overnight, the reaction mixture was cooled to 0° C. andfiltered to remove the precipitate. The filtrate was concentrated invacuo, and the residue taken up in 50 mL ethyl acetate. The ethylacetate solution was washed once with 20 mL water, once with 15 mLsaturated NaHCO₃. once with 15 mL 5% citric acid, and once with 20 mLbrine. The organic layer was dried over Na₂ SO₄, filtered andconcentrated under reduced pressure. The resulting foam was purified byradial chromatography (1.0 mm plate, 0-10% MeOH/CH₂ Cl₂ gradient),affording 487 mg (55%) of the desired product as a single diastereomeras determined by 300 MHz 1H NMR.

a!_(D) +11.93° (c 0.10, MeOH)

¹ H NMR (300 MHz, CDCL₃) d 1.46 (s, 9H, --C(CH₃)₃), 2.71 (dd, J=15, 6Hz, 1H), 2.81-3.01 (m, 4H), 3.07 (dd, J=15, 3 Hz, 1H), 3.75-3.78 (m,1H), 4.28-4.32 (m, 1H), 4.95 (dd, J=12, 6 Hz, 1H), 5.74 (br s, 1H), 6.19(br s, 1H), 6.32 (br.s, 1H), 6.90 (t, J=6 Hz, 1H), 7,01 (t, J=6 Hz, 1H),7.08-7.38 (m, 6H), 7.64 (t, J=6 Hz, 1H), 7.79 (t, J=9 Hz, 1H), 7.80 (d,J=6 Hz, 1H), 8.17-8.31 (m, 3H), 9.22 (d, J=9 Hz, 1H).

¹³ C NMR (75.4 MHz, CDCl₃) d 28.7, 35.3, 37.1, 37.2, 50.1, 52.1, 53.5,55.9, 74.8, 118.7, 126.0, 127.0, 127.6, 128.2, 129.3,129.4, 130.0,130.2, 130.3, 130.9, 137.4, 137.5, 138.2, 146.5, 148.8, 164.6, 170.4,170.5.

IR (CHCl₃) 3428, 3411, 3359, 30 12, 2975, 1681, 160 1, 1565, 1518, 1499,1454, 1428, 1395, 1367, 1231, 1047 cm⁻¹.

MS (FD) m/e 610 (M+), 221(100).

Analytical calc'd for C₃₅ H₃₉ N₅ O₅ : C 68.95, H 6.45, N 11.49; found C68.76, H 6.55, N 11.49.

EXAMPLE 2 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-4-methylphenyl)hexyl)-2-quinolinylcarboxamide

Preparation of N-t-butyl-2,5-dimethylbenzamide

The title compound was prepared according to the procedure in Example 1.Recrystallization from hexane/ethyl acetate afforded 67.2 g of thedesired product (67%), mp 80°-82° C.

¹ H NMR (300 MHz, CDCl₃) d 1.44 (s, 9H, --C(CH₃)₃), 2.29 (s, 3H), 2.36(s, 3H), 5.58-5.46 (br.s, 1H), 7.06 (s, 2H), 7.10 (s, 1H)

IR (CHCl₃) 3430, 3009, 2970, 2928, 2871, 1662, 1551, 1453, 1355, 1236cm⁻¹.

MS (FD) m/e 205(M+), 205(100).

Analytical calc'd for C₁₃ H₁₉ NO: C 76.05, H 9.33, N 6.82; found C76.30, H 9.60, N 6.83.

Preparation of(S)-N-C-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-oxopropyl)-5-methylbenzamide

The title compound was prepared according to the procedure outlined inExample 1. Flash chromatography with 20-50% ethyl acetate/hexaneprovided 6.31 g of the product ketone (89%) as a crystalline solid.

a!_(D) -25.74° (c 0.30, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.38 (s, 9H), 2.32 (s, 3H), 2.98 (dd, H =14,8 Hz, 1H), 3.22 (dd, J=14, 5 Hz, 1H), 3.93 (d, J=18 Hz, 1H), 4.10 (d,J=18 Hz, 1H), 4.65-4.76 (m, 1H), 5.00-5.10 (m, 2H), 5.58 (d, J=6.7 Hz,1H), 5.94 (s, 1H), 6.92 (d, J=8Hz, 1H), 7.09-7.39 (m, 12H).

IR (CHCl₃) 3481, 3029, 3012, 2972, 1713, 1656, 1496, 1454, 1236, 1045cm⁻¹.

MS (FD) m/e 486(M+), 486(100).

Analytical calc'd for C₃₀ H₃₄ N₂ O₄ : C 74.05, H 7.04, N 5.76; found C73.99, H 7.28, N 5.65.

Preparation of R-(R*,S*)!-N-t-butyl-2-(3-(N-phenyl-methoxy-carbonyl)amino-3-phenylmethyl-2-hydroxypropyl)-5-methylbenzamide

The title compound was prepared according to the procedure in Example 1.Flash chromatography with 2-50% ethyl acetate/hexane afforded thedesired alcohol (4.84 mg, 82%) as a colorless foam.

a!_(D) +10.31° (c 0.58, MeOH) .

¹ H NMR (300 MHz, CDCl₃) d 1.46 (s, 9H), 2.32 (s, 3H), 2.76 (d, J=15 Hz,1H), 2.85-2.95 (m, 2H), 3.04 (dd, J=15, 5 Hz, 1H), 3.67-3.74 (m, 1H),3.92-4.05 (m, 1H), 4.92-5.14 (m, 3H), 5.86 (d, J=7 Hz, 1H), 5.91 (s,1H), 7.03-7.40 (m, 13H).

IR (CHCl₃) 3431, 3274, 3027, 3012, 2970, 1713, 1643, 1514, 1496, 1454,1367, 1224, 1039, 910 cm⁻¹.

MS (FD) m/e 489 (M⁺), 205 (100).

Analytical calc'd for C₃₀ H₃₆ N₂ O₄ : C 73.74, H 7.43, N 5.73; found C73.55, H 7.50, N 5.96.

Preparation of R-(R*,S*)!-N-t-butyl-2-(3-amino-2-hydroxypropyl)-5-methylbenzamide

The title compound was prepared according to the procedure given inExample 1. Concentration of the crude filtrate afforded the aminoalcohol (4.2 g, 89%) as a colorless foam that was carried on directly tothe next step without any further purification.

Preparation of 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-4-methylphenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared as described in Example 1. Flashchromatography with 2-8% MeOH/CH₂ Cl₂ provided 2.00 g (80%) of thedesired product (>90% purity as determined by ¹ H NMR). Furtherpurification of the product by reverse phase preparatory chromatographyprovided 1.95 g (74%) of analytically pure material.

a!_(D) +28.57° (c 0.10, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.43 (s, 9H), 2.30 (s, 3H), 2.63-3.05 (m,6H), 3.65-3.76 (m, 1H), 4.22-4.35 (m, 1H), 4.89-4.97(m, 1H), 5.51 (s,1H), 5.82-5.88 (br.s, 1H), 6.09 (s, 1H), 6.20 (s, 1H), 6.84-7.24 (m,10H), 7.63 (t, J=7.5 Hz, 1H), 7.78 (t, J=7.5 Hz, 1H), 7.85 (d, J=8 Hz,1H), 8.19 (t, J=8.4 Hz, 1H), 8.28 (d, J=8.4 Hz, 1H), 9.20 (d, J=8.1 Hz,1H).

IR (CHC13) 3410, 3022, 3013, 1674, 1519, 1497, 1454, 1428, 1367, 1210,1047, 910 cm⁻¹.

MS (FD) m/e 624 (M⁺), 234 (100).

Analytical calc'd for C₃₆ H₄₁ N₅ O₅ : C 69.32, H 6.63, N 11.23; found C68.73, H 6.76, N 11.07.

EXAMPLE 3 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-3-methylphenyl)hexyl)-2-quinolinylcarboxamide

Preparation of N-t-butyl-2,6-dimethylbenzamide

The title compound was prepared according to the procedure in Example 1.Recrystallization from hexane/ethyl acetate afforded 42.3 g of thedesired product (75%), mp 134°-136° C.

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H, --C(CH₃)₃), 2.32 (s, 6H, 2 xo-CH₃), , 5.45(br.s, 1H, -NH), 6.98 (d, J=6 Hz, 2H, m-ArH), 7.12 (t,J=9Hz, 1H, p-ArH).

IR (CHCl₃) 3426, 3009, 2970, 2929, 1663, 1508, 1452, 1360, 1213 cm⁻¹.

MS (FD) m/e 205(M+), 205(100).

Analytical calc'd for C₁₃ H₁₉ NO: C 76.05, H 9.33, N 6.82; found C76.35, H 9.53, N 6.80.

Preparation of(S)-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-oxopropyl)-6-methylbenzamide

The title compound was prepared according to the procedure outlined inExample 1. Flash chromatography with 10-100% ethyl acetate/hexaneprovided 2.28 g of the product ketone (80%) as a colorless foam.

a!_(D) -17.00° (c 0.10, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.37 (2, 9H, t-Bu), 2.33 (s, 3H, o-Me), 2.98(dd, J=15, 6 Hz, 1H, PhCH₂ CH), 3.15 (dd, 15, 6 Hz, 1H, PhCH₂ CH), 3.73(d, J=15 Hz, 1H, ArCH₂ CO), 3.84 (d, J=15 Hz, 1H, ArCH₂ CO), 4.70 (dd,J=15,6 Hz, 1H ArCH₂ CH), 4.98-5.02 (m, 2H, PhCH₂ O), 5.45 (d, J=9 Hz,1H, --NHCH), 5.94 (s, 1H, --NHt-Bu), 6.81 (d, J=6 Hz, 1H), 7.06-7.31 (m,12H, ArH).

IR (CHCl₃) 3423, 3030, 3012, 2968, 1713, 1659, 1507, 1455, 1393, 1367,1305, 1229, 1218, 1045 cm⁻¹.

MS (FD) m/e 487(M+), 337(100).

Analytical calc'd for C₃₀ H₃₄ N₂ O₄ : C 74.05, H 7.04, N 5.76; found C72.44, H 6.82, N 5.54.

Preparation of R-(R*,S*)-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-hydroxypropyl)-6-methylbenzamide

The title compound was prepared according to the procedure in Example 1.Flash chromatography with 2-50% EtOAC/CH₂ Cl₂ afforded the desiredalcohol (852 mg, 85%) as a colorless foam.

a!_(D) -4.95° (c 0.10, MeOH)

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H, t-Bu), 2.35 (s, 3H, o-CH₃),2.68-2.94 (m, 4H, ArCH₂ CO, PhCH₂ CH), 3.76-3.82 (m, 1H), 3.92-4.04 (m,1H), 4.93-5.03 (m, 2H, PhCH₂ O), 5.20 (d, J=6 Hz, 1H, --NH or OH), 5.85(s, 1H), 7.04 (d, J=6 Hz, 1H), 7.19-7.31 (m, 13H, ArH).

IR (CHCl₃) 3424, 3012, 2967, 1714, 1642, 1512, 1454, 1230, 1028 910cm⁻¹.

MS (FD) m/e 489 (M⁺), 489 (100).

Analytical calc'd for C₃₀ H₃₆ N₂ O₄ : C 73.74, H 7.43, N 5.73; found C74.02, H 7.57, N 5.79.

Preparation of R-(R*,S*)!-N-t-butyl-2-(3-amino-2-hydroxypropyl)-6-methylbenzamide

The title compound was prepared according to the procedure given inExample 1. Concentration of the crude filtrate afforded the aminoalcohol (282 mg, 82%) as a colorless foam that was carried on directlyto the next step without any further purification.

Preparation of 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-3-methylphenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared as described in Example 1. Flashchromatography with 3% MeOH/CH₂ Cl₂ provided 122 mg (24%) of the desiredproduct (>95% purity as determined by ¹ H NMR).

a!_(D) +7.38° (c 0.054, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.44 (s, 9H, t-Bu), 2.32 (s, 3H, o-CH₃ ),2.64-2.96 (m, 3H, PhCH₂ CH and ArCHCO), 3.00 (dd, J=9, 6 Hz, 1H, PhCH₂CH), 3.73-3.85 (m, 1H), 4.15-4.28 (m, 1H), 4.83-4.97 (m, 1H), 5.62 (s,1H), 6.10 (s, 1H), 6.29 (s, 1H), 6.86-7.25 (m, 7H, ArH), 7.62 (t, J=6Hz, 1H), 7.77 (t, J=6 Hz, 1H), 7.86 (d, J=9 Hz, 1H), 8.13-8.29 (m, 2H),9.14 (d, J=6 Hz, 1H).

IR (CHCl₃) 3413, 3017, 1681,1595, 1520, 1500, 1455, 1428, 1368, 1209,910 cm⁻¹.

MS (FD) m/e 624 (M⁺), 624 (100).

Analytical calc'd for C₃₆ H₄₁ N₅ O₅ : C 69.32, H 6.63, N 11.23; found C68.73, H 6.76, N 11.07.

EXAMPLE 4 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(1-(1-t-butylamino-1-oxomethyl)-2-naphthyl)hexyl)-2-quinolinylcarboxamide

Preparation of N-t-butyl-2-methyl-1-naphthylamide

The title compound was prepared according to the procedure given inExample 1. Recrystallization from hexane/ethyl acetate afforded 20.99 g(68%) of the desired amide as colorless needles, mp 124°-126° C. NMR(300 MHz, CDCl₃) d 1.54 (s, 9H, t-Bu), 2.50 (s, 3H), 5.50-5.65 (br.s,1H), 7.23-7.54 (m, 3H), 7.74 (d, J=10 Hz, 1H), 7.78 (d, J=10 Hz, 1H),7.87 (d, J=10 Hz, 1H).

IR (CHCl₃) 3424, 3010, 2969, 1660, 1512, 1503, 1454, 1366, 1291, 1263,1221 cm⁻¹.

MS (FD) m/e 241(M⁺), 241(100).

Analytical calc'd for C₁₆ H₁₉ NO: C 79.63, H 7.94, N 5.80; found C79.90, H 8.11, N 5.76.

Preparation of(S)-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-oxopropyl)-1-naphthylamide

The title compound was prepared according to the procedure in Example 1.Flash chromatography with 10-30% ethyl acetate/hexane provided 7.43 g(86%) of the desired ketone as a colorless foam.

a!_(D) -6.86° (c 0.10, MeOH).

NMR (300 MHz, CDCl₃) d 1.45 (s, 9H, t-Bu), 3.03 (dd, J=15, 8 Hz, 1H),3.18 (dd, J=15, 5 Hz, 1H), 3.91 (d, J=16 Hz, 1H), 4.04 (d, J=16 Hz, 1H),4.70-4.80 (m, 1H), 4.94-5.06 (m, 2H), 5.41 (d, J=8 Hz, 1H), 6.12-6.20(br.s, 1H), 7.10-7.38 (m, 11H), 7.42-7.58 (m, 2H), 7.76-7.85 (m, 2H),7.93 (s, J=9 Hz, 1H).

IR (CHCl₃) 3420, 3029, 3012, 2970, 1713, 1658, 1505, 1455, 1367, 1232,1045 cm⁻¹.

MS (FD) m/e 522(M⁺), 522(100).

Analytical calc'd for C₃₃ H₃₄ N₂ O₄ : C 75.84, H 6.56, N 5.36; found C75.56, H 6.74, N 5.17.

Preparation of R-(R*,S*)!-N-t-butyl-2-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-hydroxypropyl)-1-naphthylamide

The title compound was prepared according to the procedure in Example 1.Flash chromatography with 2-10% ethyl acetate/dichloromethane provided5.50 g (74%) of the desired alcohol diastereoisomer as a colorless foam.

a!_(D) +11.85° (c 0.20, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.54 (s, 9H, t-Bu), 2.85-3.15 (m, 4H),3.85-3.95 (m, 1H), 4.00-4.13 (m, 2H), 4.90-5.34 (m, 3H), 5.85-5.95 (m,1H), 7.05-7.60 (m, 15H), 7.81 (d, J=9 Hz, 2H), 7.91 (d, 9 Hz, 2H).

IR (CHCl₃) 3420, 3012, 2970, 1713, 1643, 1515, 1454, 1367, 1219, 1209,1028 cm⁻¹.

MS (FD) m/e 524(M+), 524(100).

Analytical calc'd for C₃₃ H₃₆ N₂ O₄ : C 75.55, H 6.92, N 5.34; found C75.41, H 7.16, N 5.14.

Preparation of R-(R*,S*)!-N-t-butyl-2-(3-amino-2-hydroxypropyl)-1-naphthylamide

The title compound was prepared according to the procedure given inExample 1. Concentration of the crude filtrate afforded the aminoalcohol (1.30 g, 92%) as a colorless foam that was carried on directlyto the next step without any further purification.

Preparation of 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(1-(1-t-butylamino-1-oxomethyl)-2-naphthyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared as described in Example 1. Flashchromatography with 2-5% MeOH/CH₂ Cl₂ provided 1.75 g (80%) of thedesired product (>90% purity as determined by ¹ H NMR). Furtherpurification of the product by reverse phase preparatory chromatographyprovided 1.06 g (48%) of analytically pure material.

a!_(D) +18.82° (c 0.15, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.46 (s, 9H), 2.42-2.80 (m, 4H), 3.00 (br.d,J=13.5 Hz, 2H), 3.60-3.93 (m, 2H (rotamers)), 4.65-4.75 (m, 1H), 5.05(br.s, 0.4H), 5.38 (br.s, 0.6 H), 6.85-6.93 (m, 2H), 7.04 (t, J=7.5 Hz,2H), 7.18 (d, J=7.4 Hz, 2H), 7.30-7.54 (m, 4H), 7.70 (t, J=7.5 Hz, 1 H),7.78-7.90 (m, 3H), 7.95-8.12 (m, 3H), 8.17 (d, J=8.5 Hz, 1H), 8.56 (d,J=8.5 Hz, 1H), 8.84 (d, J=7.6 Hz, 1H).

IR (CHCl₃) 3414, 3010, 1680, 1519, 1499, 1454, 1428, 1367, 213, 846cm⁻¹.

MS (FD) m/e 660 (M⁺), 660 (100).

EXAMPLE 5 1S-(1R*. 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(3-(1-t-butylamino-1-oxomethyl)-2-naphthyl)hexyl)-2-quinolinylcarboxamide

Preparation of N-t-butyl-3-methyl-2-naphthylamide

The title compound was prepared according to the procedure given inExample 1. Recrystallization from hexanes afforded 6.80 g (80%) of thedesired product, mp 95°-96° C.

¹ H NMR (300 MHz, CDCl₃) d 1.50 (s, 9H), 2.56 (s, 3H), 5.70 (br.s, 1H),7.40-7.5 (m, 2H), 7.62 (s,: 1H), 7.72-7.80 (m, 3H).

IR (CHCl₃) 3430, 3011, 2970, 1666, 1512, 1496, 1452, 1366, 212, 884cm⁻¹.

MS (FD) m/e 241(M⁺), 241 (100).

Analytical calc'd for C₁₆ H₁₉ NO: C 79.63, H 7.94, N 5.80; found C79.57, H 8.19, N 5.71.

Preparation of(S)-N-t-butyl-3-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-oxopropyl)-2-naphthylamide

The title compound was prepared according to the general procedure givenin Example 1. Flash chromatography with 3:1 hexanes/ethyl acetate gave3.41 g (79%) of the desired product as a colorless oil.

a!_(D) -32.10° (c 1,028, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.43 (s, 9H), 3.01 (dd, J=13.8, 7.3 Hz, 1H),3.27 (dd, J=14.90, 6.03 Hz, 1H), 4.06 (d, J=18.3 Hz, 1H), 4.34 (d,J=17.4 Hz, 1H), 4.72-4.79 (m, ¹ H), 4.98-5.09 (m, 1H), 5.59 (br.d, J=7.8Hz, 1H), 6.02 (s, 1H), 7.19-7.35 (m, 10H), 7.48-7.51 (m, 3H), 7.75 (d,J=8.15 Hz, 1H), 7.81 (d, J=8.9 Hz, 1H), 7.87 (s, 1H).

IR (CHCl₃) 3400, 3011, 2970, 1713, 1657, 1512, 1454, 1301, 1234, 1105cm⁻¹.

MS (FD) m/e 523(M+), 523(100).

Analytical calc'd for C₃₃ H₃₄ N₂ O₄ : C 75.84, H 6.56, N 5.36; found C75.75, H 6.57, N 5.33.

Preparation of R-(R*,S*)!-N-t-butyl-3-(3-(N-phenylmethoxycarbonyl)amino-3-phenylmethyl-2-hydroxypropyl)-2-naphthylamide

The title compound was prepared according to the procedure outlined inExample 1. Flash chromatography with 19:1 hexanes/ethyl acetate afforded1.87 g (59%) of the desired compound as a colorless foam.

a!_(D) +11.02° (c 0.998, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.53 (s, 9H), 2.93-3.04 (br.m, 5H), 3.85-3.89(m, 1H), 4.07-4.12 (m, 1H), 5.00-5.10 (m, 2H), 5.16 (d, J=8.8 Hz, 1H),6.19 (s, 1H), 7.22-7.40 (m, 10H), 7.48-7.56 (m, 2H), 7.74-7.87 (m, 4H).

IR (CHCl₃) 3431, 3027, 3011, 1713, 1645, 1514, 1454, 1367, 1223, 1044cm⁻¹.

MS (FD) m/e 524 (M⁺), 270 (100).

Preparation of 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(3-(1-t-butylamino-1-oxomethyl)-2-naphthyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedure inExample 1. Radial chromatography (1 mm silica plate, 0-10% MeOH/CH₂ C₁₂)afforded 162 mg of the product (74%) as a colorless foam.

a!_(D) +22.83° (c 0.876, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.52 (s, 9H), 2.74 (dd, J=15.1, 8.3 Hz, 1H),2.90-2.98 (m, 3H), 3.10 (apparent dd, J=14.8, 9.8 Hz, 3H), 3.83-3.85 (m,1H), 4.35-4.38 (m, 1H), 4.96-5.01 m, 1H), 5.47 (s, 1H), 6.09 (s, 1H)6.22 (s, 1H), 6.91-6.94 (m, 1H), 7.02 (7, J=7.6 Hz, 2H), 7.08-7.13 (m,1H), 7.22-7.27 ° (m, 3H), 7.44-7.53 (m, 2H), 7.68 (t, J=7.5 Hz, 1H),7.77-7.86 (m, 4H), 7.92 (d, J=8.0 Hz, 1H), 8.26 (d, J=8.4 Hz, 2H), 8.36(d, J=8.5 Hz, 1H), 9.32 (d, J=8.2 Hz, 1H).

IR (KBr pellet) 3319, 1659, 1522, 1497, 1427, 1220, 911, 846, 733, 477cm⁻¹.

MS (FD) m/e 661 (M⁺), 661 (100).

Analytical calc'd for C₃₉ H₄₁ N₅ O₅ : C 71.00, H 6.26, N 10.61; found C71.24, H 6.15, N 10.51.

By following the procedures described in Example 1 and employing theappropriate reactants, the compounds of Examples 6 through 27 wereprepared.

EXAMPLE 6 1S-(1R*, 4R*,5R*)!-N-(1-(2-amino-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 36% yield. ¹ H NMR (300 MHz, CDCl₃) d1.41 (s, 9H), 2.69-3.07 (m, 7H), 3.77-3.84 (m, 1H), 4.23-4.34 (m, 1H),4.91-5.03 (m, 1H), 5.52-5.62 (m, 1H), 5.98 (s, 1H), 6.32 (br.s, 1H),7.00 (d, H=8Hz, 1H), 7.06-7.34 (m, 9H), 7.62 (t, J=7.5 Hz, 1H), 7.77 (t,J=7.5 Hz, 1H), 7.87 (d, J=7.5 Hz, 1H), 8.17-8.32 (m, 3H), 9.20 (d, J=8Hz, 1H).

IR (CHCl₃) 3012, 1681, 1519, 1499, 1222 cm⁻¹.

MS (FD) m/e 610 (M⁺), 610 (100).

Analytical calc'd for C₃₅ H₃₉ N₅ O₅ : C 68.94, H 6.45, N 11.49; found C68.37, H 6.45, N 11.00.

EXAMPLE 7 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-1-oxo-2-quinolinylcarboxamide

The title compound was isolated in 23% yield.

a!_(D) +34.03° (c 0.529, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.44 (s, 9H), 2.71-2.97 (m, 5H), 3.07 (dd,J=14.3, 4.4 Hz, 1H), 3.73-3.76 (m, 1H), 4.26-4.29 (m, 1H), 4.97-4.99 (m,1H), 5.72 (s, 1H), 5.91 (br.s, 1H), 6.28 (s, 1H), 6.33 (s, 1H), 6.84 (t,J=7.3 Hz, 1H), 7.00 (t, J=7.5 Hz, 2H), 7.09-7.33 (m, 7H), 7.67-7.87 (m,4H), 8.21 (d, J=8.9 Hz, 1H), 8.77 (d, J=8.7 Hz, 1H), 12.05 (d, J=7.6 Hz,1H).

IR (KBr) 3311, 3065, 2966, 1656, 1520, 1453, 1366, 735, 701 cm⁻¹.

Analytical calc'd for C₃₅ H₃₉ N₅ O₆ : C 67.18, H 6.26, N 11.19; found C66.92, H 6.32, N 11.21.

EXAMPLE 8 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-(1.1-dimethylpropylamino)-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 34% yield.

a!_(D) +23.60° (c 0.10, MeOH)

¹ H NMR (300 MHz, CDCl₃) d 0.89 (t, J=7 Hz, 3H), 1.38 (s, 6H), 1.81 (q,J=7 Hz, 2H), 2.69 (dd, J=15, 7 Hz, 1H), 2.79-2.99 (m, 4H), 3.04 (dd,J=14, 4 Hz, 1H), 3.71-3.75 (m, 1H), 4.26-4.30 (m, 1H), 4.91-4.96 (m,1H), 5.61 (s, 1H), 5.98 (s, 1H), 6.26 (s, 1H), 6.86-7.37 (m, 11H), 7.62(t, J=7.5 Hz, 1H), 7.77 (t, J=7.5 Hz, 1H), 7.87 (d, J=8.2 Hz, 1H),8.16-8.30 (m, 3H), 9.20 (d, J=8.0 Hz, 1H).

IR: (CHCl₃) 3410, 3028, 3010, 1681, 1518, 1499, 1428, 1234, 1048, 846cm⁻¹.

MS (FD) 624(M⁺) 624(100)

Analytical calc'd for C₃₆ H₄₁ N₅ O₅ : C 69.32, H 6.63, N 11.23; found C69.06, H 6.76, N 10.98.

EXAMPLE 9 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-(4-phenylmethyloxy)phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 67% yield.

a!_(D) 0.00° (c 2.0, MeOH); a!₃₆₅ +76.24° (c 2.0, MeOH) ¹ H NMR (300MHz, CDCl₃) d 1.44 (s, 9H), 2.72 (dd, J=15.3, 7.0 Hz, 1H), 2.81-2.99 (m,4H), 3.08 (q, J=7.3 Hz, 1H), 3.67-3.71 (m, 1H), 4.54-4.63 (m, 2H),4.93-4.99 (m, 1H), 5.54-5.59 (br s, 1H), 5.90 (s, 1H), 6.08-6.16 (br.s,1H), 6.56 (d, J=8, 4 Hz, 2H), 7.06-7.13 (m, 3H), 7.25-7.37 (m, 10H),7.59 (t , J=7.5 Hz, 1H), 7.73 (t, J=7.3 Hz, 1H), 7.84 (d, J=8.2 Hz, 1H),8.17-8.30 (m, 3H), 9.26 (d, J=8.2 Hz, 1H).

IR (CHCl₃) 3410, 3025, 3011, 1680, 1512, 1500, 1454, 1367, 1218 cm⁻¹.

MS (FD) m/e 716 (M⁺), 716 (100).

Analytical calc'd for C₄₂ H₄₅ N₅ O₆ : C 70.47, H 6.34, N 9.78; found C70.65, H 6.43, N 9.74.

EXAMPLE 10 1S-(1R*, 4R*. 5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-(4-hydroxyphenyl)methyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 87% yield.

a!_(D) +17.43° (c 1.0, MeOH)

¹ H NMR (300 MHz, CDCl₃) d 1.42 (s, 9H), 2.62-2.76 (m, 2H), 2.84-3.02(m, 3H), 3.74-3.78 (m, 1H), 4.03-4.06 (m, 1H), 4.88-4.92 (m, 1H), 6.45(d, J=8.3 Hz, 2H), 6.98 (d, J=8.3 Hz, 2H), 7.20-7.37 (m, 4H), 7.67 (t,J=7.5 Hz, 1H), 7.82 (t, J=7.8 Hz, 1H), 7.78-7.80 (m, 2H), 8.17 (d, J=8.1Hz, 1H), 8.47 (s, J=8.5 Hz, 1H).

IR (KBr) 3308, 2967, 2341, 1658, 1516, 1499, 1226, 775, 733 cm⁻¹.

MS (FD) m/e 626 (M⁺), 626 (100)

EXAMPLE 11 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-(2-amino-2-oxomethyl)-6-phenylmethoxycarbonyl)hexyl)benzamide

The title compound was isolated in 52% yield.

a!_(D) -6.72° (c 1.042, MeOH).

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H), 2.77-3.08 (m, 7H), 3.49 (d,J=11.1, 5.42 Hz, 1H), 3.76-3.86 (m, 1H), 4.09 (s, 1H), 4.30-4.46 (m,1H), 5.06 (s, 2H), 5.64 (d, J=6.9 Hz, 1H), 6.04 (s, 1H), 6.80 (d, J=8.8Hz, 1H), 7.16-7.46 (m, 14H).

IR (CHCl3) 3011, 2974, 1716, 1673, 1641, 1419, 1050 cm⁻¹.

MS (FD) m/e 589 (M⁺), 589 (100) .

Analytical calc'd for C₃₃ H₄₀ N₃ O₆ : C 67.33, H 6.85, N 9.52; found C67.21, H 6.77, N 9.41.

EXAMPLE 12 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-hydroxymethyl-6-(N-phenylmethoxycarbonyl-amino)hexyl)benzamide

The title compound was isolated in 22% yield.

a!_(D) +2.58 (c 0.388, MeOH).

¹ H NMR (300 MHz, CD₃ OD) d 1.43 (s, 9H), 2.40 (dd, J=14.0, 7.7 Hz, 1H),2.56 (dd, J=15.5, 6.1 Hz, 1H), 2.74 (dd, J=13.5, 9.9 Hz, 1H), 2.83-2.96(m, 2H), 3.11 (dd, J=-14.0, 3.5 Hz, 1H), 3.72-3.79 (m, 1H), 4.02-4.08(m, 1H), 4.41 (dd, J=7.6, 6.4 Hz, 1H), 5.06 (s, 2H), 7.09-7.38 (m, 14H).

IR (KBr) 3305, 3065, 2966, 1718, 1638, 1535, 1454, 1326, 1220, 1060cm⁻¹.

MS (FD) m/e 562 (M⁺), 220 (100).

EXAMPLE 13 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-1-quinolinyl-2-carboxamide

The title compound was isolated in 22% yield.

a!_(D) -9.71° (c 0.206, MeOH)

¹ H NMR (300 MHz, CDCl³) d 1.42 (s, 9H), 2.33 (br. s, 1H), 2.65 (dd,J=15.2, 6.7 Hz, 1H), 2.79-2.97 (m, 14H), 3.03 (dd, J=14.0, 3.8 Hz, 1H),3.72 (br.m, 1H), 4.23 (br.m, 1H), 4.92 (br.m, 1H), 5.71 (br.s, 1H), 6.27(s, 1H), 6.44 (br.s, 1H), 6.86-6.98 (m, 2H), 7.12-7.31 (m, 7H),7.59-7.71 (m, 2H), 7.77 (d, J=5.4 Hz, 1H), 7.82 (d, J=7.94 Hz, 1H), 8.46(d, J=5.5 Hz, 1H), 8.47 (d, J=8.0 Hz, 1H), 9.41 (d, J=8.3 Hz, 1H).

IR (CHC13) 3010, 2974, 1680, 1601, 1515, 1454, 1394, 1230 cm⁻¹.

MS (FD), m/e 610 (M+) , 220 (100).

EXAMPLE 14 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-benzimidazolylcarboxamide

The title compound was isolated in 63% yield.

a!_(D) -16.85° (c 0.53, MeOH).

¹ H NMR (300 MHz, CD₃ OD) d 1.43 (s, 9H), 2.59-2.77 (m, 4H), 2.93-3.00(m, 2H), 3.14 (dd, J=12, 4 Hz, 1H), 3.72-3.83 (m, 1H), 4.06-4.13 (m,1H), 6.81 (t, J=7 Hz, 1H), 6.97 (t, J=7.5 Hz, 2H), 7.14-7.38 (m, 8H),7.58-7.64 (br.m, 2H).

IR (KBr) 3318, 3062, 2965, 2405, 1639, 1546, 1445, 1422, 1392, 1228cm⁻¹.

MS (FD) m/e 599(M+), 599(100).

Analytical calc'd for C₃₃ H₃₈ N₆ O₅ : C 6.20, H 6.40, N 14.04; found C66.14, H 6.56, N 13.78.

EXAMPLE 15 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4,7-diaza-5,8,-dioxo-6-(2-amino-2-oxoethyl)-9-(naphthyloxy)nonyl)benzamide

The title compound was isolated in 62% yield.

a!_(D) -2.00° (c 1.0, MeOH)

¹ H NMR (300 MHz, CD₃ OD) d 1.43 (s, 9H), 2.59-2.98 (m, 6H), 3.09 (br.d,J=3.9 Hz, 1H), 3.72-3.82 (m, 1H), 4.03-4.14 (m, 1H), 4.64-4.79 (m, 2H),6.85 (d, J=7.5 Hz, 1H), 6.97 (t, J=2.0 Hz, 1H), 7.10-7.52 (m, 12H),7.79-7.82 (m, 1H), 8.34-8.36 (m, 1H).

IR (KBr) 3296, 3061, 2967, 2930, 2540, 2413, 1646, 1427, 1401, 1236, 768cm⁻¹.

MS (FD) m/e 639(M⁺), 639(100).

EXAMPLE 16 1S-(1R*, 4R*,5S*)!-N-(1-(2-propyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 13% yield.

¹ H NMR (300 MHz, CDCl₃) d 0.87 (d, J=6.6 Hz, 3H), 0.97 (d, J=6.6 Hz,3H), 1.46 (s, 9H), 2.30-2.37 (m, 1H), 2.79-3.09 (m, 4H), 3.72-3.80 (m,1H), 4.31-4.41 (m, 2H), 5.96 (s, 1H), 6.04-6.10 (m, 1H), 6.49 (d, J=9.1Hz, 1H), 6.80-6.89 (m, 1H), 7.04 (t, J=7.5 Hz, 1H), 7.20-7.40 (m, 7H),7.64 (t, J=7.5 Hz, 1H), 7.80 (t, J=7.6 Hz, 1H), 7.90 (d, J=8.1 Hz, 1H),8.16 (d, J=8.4 Hz, 1H), 8.30 (q, J=8.5 Hz, 2H), 8.53 (d, J=9.0 Hz, 1H).

MS (FAB) m/e 595 (M⁺), 128 (100).

EXAMPLE 17 1S-(1R*, 4R*.5S*)!-N-(1-(4-imidazolylmethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 84% yield.

a!_(D) -8.46° (c 1.06, MeOH)

¹ H NMR (300 MHz, CDCl₃) d 1.49 (s, 9H), 2.75-2.86 (m, 2H), 2.98-3.18(m, 2H), 3.21 (d, J=6.5 Hz, 2H), 3.93-3.99 (m, 2H), 4.32-4.43 (m, 1H),4.81-4.86 (m, 1H), 6.50 (br.s, 1H), 8.84 (s, 1H), 6.95-7.42 (m, 9H),7.58-7.65 (m, 1H), 7.75-7.79 (m, 1H), 7.86 (d, J=7.5 Hz, 1H), 8.10 (d,J=8 Hz, 1H), 8.19 (d, J=7.5 Hz, 1H), 8.25 (d, J=7.5 Hz, 1H), 8.88 (d,J=8 Hz, 1H).

IR (KBr) 3700, 3276, 3010, 1656, 1601, 1518, 1500, 1455, 428, 1367,1231, 1095, 830 cm⁻¹.

MS (FD) m/e 633 (M⁺), 633 (100) .

Analytical calc'd for C₃₇ H₄₀ N₆ O₄ : C 70.23, H 6.37, N 13.28; found C70.26, H 6.41, N 13.23.

EXAMPLE 18 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(2,2-dimethyl-1-oxopropylamino)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was isolated in 61% yield.

a!_(D) -53.42° (c 1.003, MeOH)

¹ H NMR (300 MHz, CDCl₃) d 1.31 (s, 9H), 2.62-3.03 (m, 6H), 4.06(apparent dd, J=9.9 Hz, 1H), 4.17-4.22 (m, 1H), 4.85-4.91 (m, 1H), 5.52(s, 1H), 6.00 (s, 1H), 6.94 (t, J=7.4 Hz, 1H), 7.02-7.27 (m, 9H), 7.64(t, J=7.1 Hz, 1H), 7.77 -7.81 (m, 2H), 7.86 (d, J=8.1 Hz, 1H), 8.14 (d,J=8.4 Hz, 2H), 8.27 (d, J=8.5 Hz, 1H), 9.14 (d, J=7.7 Hz, 1H), 9.23 (s,1H).

IR (KBr) 3329, 1659, 1498, 1450, 845, 753 cm⁻¹.

MS (FD) m/e 610 (M⁺), 610 (100).

Analytical calc'd for C₃₅ H₃₉ N₅ O₅ : C 68.95, H 6.45, N 11.49; found C69.12, H 6.57, N 11.57.

EXAMPLE 19 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-naphthylcarboxamide

Yield: 30 mg (12%) as an off-white foam

¹ H NMR (CDCl₃) d 8.33 (s, 1H,), 8.15 (d, 1H), 8.0-6.95 (m, 15H,), 5.98(br.s, 2H), 5.37 (br.s, 1H), 4.89 (m, 1H), 4.3 (m, 1H), 3.78 (m, 1H),3.1-2.58 (m, 6H), 1.45 (s, 9H). MS (FD) 609 (P+1).

EXAMPLE 20 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinoxalinylcarboxamide

Yield: 29 mg (7%) of a yellowish foam

¹ H NMR (CDCI₃) d 9.52 (s, 1H), 9.03 (d, 1H), 8.15-6.18 (m, 13H), 6.31(br.s, 1H), 6.15 (s, 1H), 5.92 (d, 1H), 5.7 (br.s, 1H), 4.91 (m, 1H),4.3 (m, 1H), 3.77 (m, 1H), 3.07-2.6 (m, 6H), 1.41 (s, 9H).

MS (FD) 612 (P+2), 593, 519, 391, 307, 243, 220.

Analytical calc'd for: C₃₄ H₃₈ N₆ O₅ : C 66.87, H 6.27, N 13.76; found:C 67.14, H 6.49, N 13.56.

EXAMPLE 21 1S-(1R*, 4R*.5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-indolylcarboxamine

Yield: 86 mg (16%) of a slightly colored foam.

¹ N HMR (DMSO-d₆) d 11.6 (3, 1H), 8.43 (d, 1H), 8.2 (s, 1H), 7.8 (d,1H), 7.6-7.0 (m, 13H), 5.83 (d, 1H), 4.79 (m, 1H), 3.81 (m, 1H), 3.6 (m,1H), 3.0-2.45 (m, 6H), 1.38 (s, 9H). MS (FD) 599 (P+2).

Analytical calc'd for C₃₄ H₃₉ N₅ O₅ : C 68.32, H 6.58, N 11.71; found: C67.69, H 6.89, N 10.42.

EXAMPLE 22 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-benzob!thienyl carboxamide

Yield: 31 mg (11%) as a colorless foam.

¹ H NMR (CDCl₃) d 8.17 (d, 1H), 7.85-7.0 (m, 14H), 6.03 (d, 2H), 5.41(s, 1H), 4.82 (m, 1H), 4.3 (m, 1H), 3.78 (m, 1H), 3.1-2.6 (m, 6H), 1.5(s, 9H). MS (FD) 616 (P12).

EXAMPLE 23 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-7-methylquinolin-2-ylcarboxamide

a. 2-Carboxy-7-methylquinoline

By substantially following the procedures described in Shuman et al., J.Org. Chem., 55, 738-741 (1990), 2-carboxy-7-methylquinoline wasprepared.

b. R-(R*, S*)!-N-t-butyl-2-(3-amino-2-hydroxypropyl) benzamide

By substantially following the procedures described in Example 1, stepsa through d, the title compound was prepared.

c. R-(R*, S*, S*)!-N-t-butyl-2-(3-N-(benzyloxycarbonylasparginyl)amino)-2-hydroxypropyl) benzamide

To 20 ml of dry dimethyl formamide (DMF) was added R-(R*,S*)!-N-t-butyl-2-(3-amino-2-bydroxypropyl) benzamide (3.4 g; 10 mmol)from step b above; benzyloxycarbonyl-L-asparagine (2.6 g; 10 mmol);N-methylmorpholine (1.09 ml; 10 mmol); and 1-hydroxybenzotriazolehydrate (1.48 g; 11 mmol). After the solution was cooled to 0° C.,1,3-dicyclohexylcarbodiimide (2.4 g; 11 mmol) was added. The reactionmixture was allowed to warm to room temperature while stirring under anitrogen atmosphere. After stirring overnight, the reaction mixture wasfiltered and the filtrate extracted with ethyl acetate. To the ethylacetate solution was added an ethyl acetate/water mixture and theorganics separated. The organics were washed sequentially with NaHCO₃,5% citric acid, saturated NaHCO₃ and brine, then dried over MgSO₄. Thematerial was evaporated to dryness, rinsed with an ether/hexane mixtureand the solids filter dried in a vacuum desiccator to afford 4.4 g(78.8%) of the title compound.

d. R-(R*, S*,S*)!-N-t-butyl-2-(3-(N-aspariginylamino)-2-hydroxypropyl)benzamide

By substantially following the procedures described in Example 1, stepd, the title compound was prepared.

e. 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-7-methylquinolin-2-ylcarboxamide

To 15 ml of dry CH₂ Cl₂ was added 2-carboxy-7-methyl quinoline (135 mg;0.5 mmol) and 1,1-carbonyldiimidazole (81 mg; 0.5 mmol) and the mixturestirred at room temperature for 30 mintues. To the solution was addedthe R-(R*, S*,S*)!-N-t-butyl-2-(3-(N-aspariginylamino)-2-hydroxypropyl)benzamide (227mg; 0.5 mmol) from step d, above. The reaction mixture was stirred atroom temperature overnight. To the reaction mixture was added CH₂ Cl₂(100 ml) and 25 ml of water to afford a poor emulsion. The emulsion waswashed sequentially with 25 ml of saturated NaHCO₃ ; 25 ml of 5% citricacid; 25 ml of saturated NaHCO₃ ; and brine. The resulting material wasdried over MgSO₄ and evaporated under vacuum to dryness. The residue wasslurried with ether and the resulting precipitate filtered off and driedin a vacuum desiccator at room temperature to afford 115 mg (37% yield)of the title compound.

MS: FAB(M+1) 623+1

H¹ NMR (300 MHz, CDCl₃) d 1.46 (s, 9H), 2.6-3.1 (m, 3H₂, 1H, 1CH₃),3.69-3.75 (m, 1H), 4.22-4.32 (m, 1H), 4.28-4.35(m, 1H), 4.9 (m, 1H),5.3-5.35 (m, 1H), 5.9-6.05 (2 br.s, 2H), 6.89-8.3 (m, 14H), 9.3 (d, 1H).

By following the procedures described in Example 28 and employing theappropriate reactants, the compounds of Examples 29 through 35 wereprepared.

EXAMPLE 24 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-8-methylquinolin-2-ylcarboxamide

Yield: 57.8%

MS: FAB--M+1=623+1

¹ H NMR (300 MHz, CDCl₃) d 1.46 (s 9H); 2.6-3.1 (m, 3H₂, 1H, 1CH--),3.70-3.78, (m, 1H), 4.28-4.32 (m, 1H), 4.9 (m, 1H), 5.2 (br.s, 1H),6.05, (br.s, 2H), 6.8-7.78, (m, 12H), 8.2-8.3 (dd, 2H).

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C             69.32    69.10                                                  H             6.62     6.81                                                   N             11.23    10.94                                                  ______________________________________                                    

EXAMPLE 25 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-6-methylquinolin-2-ylcarboxamide

DCC (Dicyclahoxylcarbodiimide) was used as the coupling agent for thelast reaction.

Yield: 54.6%

MS FAB (M+1) 623+1

H¹ NMR (300 MHz, CDCl₃) d 1.46, (s, 9H), 2.57-2.6 (s, 3H), 2.62-3.1 (m,7H), 3.7-3.78, (m, 1H), 4.23-4.35 (m, 1H), 4.9-4.95 (m, 1H), 5.41-4.43(br.s, 1H), 5.8-6.1 (2 br.s, 3H), 6.82-8.2 (m, 14H), 9.18-9.2 (d, 1H).

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C             69.32    69.08                                                  H             6.62     6.75                                                   N             11.23    10.94                                                  ______________________________________                                    

EXAMPLE 26 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-4-methylquinolin-2-ylcarboxamide

Yield: 35%

MS FAB (M+1) =623+1

¹ H NMR (300 MHz, CDCl₃) d 1.46, (s, 9H), 2.6-3.1 (m, 10H),3.7-3.78 (m,1H), 4.23-4.35 (m, 1H), 4.9-4.95 (m, 1H), 5.35-5.40 (br.s, 1H),5.97-6.08 (2 br.s, 3H), 6.82-8.2 (m, 14H), 9.18-9.2 (d, 1H).

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C             69.32    69.06                                                  H             6.62     6.64                                                   N             11.23    11.25                                                  ______________________________________                                    

EXAMPLE 27 1S-(1R*, 4R*. 5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-3-methylquinolin-2-yl carboxamide

Yield: 37%

MS FAB =correct mw of M+1=623+1.

H¹ NMR (300 MHz, CDCl₃) d 1.46 (s, 9H), 2.85-3.1 (m, 10H), 3.7-3.78, (m,1H), 4.23-4.35 (m, 1H), 4.9-4.95 (m, 1H), 5.35-5.4 (br.s, 1H),5.97-6.08, (2 br.s, 3H); 6.82-8.2 (m, 14H) , 9.18-9.2 (d, 1H)

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C             69.32    68.56                                                  H             6.63     6.67                                                   N             11.23    10.48                                                  ______________________________________                                    

EXAMPLE 28 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)phenylcarboxamide

Yield: 41.8%

H¹ NMR (300 MHz, CDCl₃) d 1.46 (s, 9H), 2.48-3.1 (m, 7H), 3.68-3.79 (m,1H), 4.23-4.35 (m, 1H), 4.9-4.95 (m, 1H), 5.32-5.39, (br.s, 1H),5.98-6.07, (br.s, 2H) 7.08-7.8 (m, 1H), (8-8.03, (d, 1H).

MS (FAB) =M+1=558+1

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C             68.80    68.56                                                  H             6.86     6.92                                                   N             10.03    10.00                                                  ______________________________________                                    

EXAMPLE 29 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-pyridylcarboxamide

The final coupling reaction was accomplished using a pentafluroactivatedester prepared substantially according to the procedures described inPreparations 2 and 3.

MS FAB =M+1=559+1

¹ H NMR (300MHz, CDCl₃) d 1.46 (s, 9H), 2.59-3.1 (m, 7H), 3.68-3.79 (m,1H), 4.23-4.35 (m, 1H), 4.85-4.92 (m, 1H), 5.39-5.43, (br.s, 1H),5.99-6.07, (2 br.s, 3H), 6.98-7.5 (m, 10H), 7.8-7.88 (t, 1H), 8.1-8.17(d, 1H), 8.59-8.63 (d, 1H), 9-9.05 (d, 1H).

EXAMPLE 30 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-3-isoquinolinylcarboxamide

The final coupling reaction was carried out using an activatedpentafluoro ester prepared according to the procedures described inPreparations 2 and 3. Yield 54.7%

MS =FAB =M+1=609+1

¹ H NMR (300MHz, CDCl₃) d 1.46 (s, 9H), 2.65-3.1 (m, 7H) 3.67-3.75 (m,1H), 4.2-4.35 (m, 1H), 4.91-4.97 (m, 1H), 5.3-5.38, (br.s, 1H),5.8-6.03, (3 br.s, 4H), 6.9-8.1 (m, 12H), 8.3-8.39 (s, 1H), 9.19-9.13(m, 2H).

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C             68.95    68.68                                                  H             6.45     6.48                                                   N             11.49    11.27                                                  ______________________________________                                    

EXAMPLE 31 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-naphthylacetamide

Yield: 85 mg (62%) as a colorless foam.

¹ H NMR (300 MHz, CDCl₃) d 7.82-7.05 (m, 16H), 6.93 (d, 1H), 6.05 (m,zH), 5.3 (br.s, 1H), 4.63 (m, 1H) 4.2 (m, 1H), 3.7 (s, 2H), 3.62 (m,1H), 3.01-2.4 (m, 6H), 1.43 (s, 9H).

MS (FD) 623 (P+1).

EXAMPLE 32 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinoxyacetamide

Yield: 90 mg (64%) of a colorless foam.

¹ H NMR (300 MHZ, CDCl₃) d 8.1 (d, 1H), 7.83-6.98 (m, 16H), 6.1 (br.m,2H), 5.41 (m, 1H), 4.79 (m, 1H), 4.56 (d, 2H), 4.25 (m, 1H), 3.71 (m,1H), 3.02-2.5 (m, 6H), 1.43 (s, 9H). MS (FD) 640 (P+1).

EXAMPLE 33 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2(6,7, 8, 9-tetrahydronaphthyl) carboxamide

Yield: 75 mg (54%) as a colorless foam.

¹ H NMR (300 MHz, CDCl₃) d 8.0 (d, 1H), 7.5-7.03 (m, 12H), 6.51 (br.s,1H), 6.22 (br.s, 1H), 5.57 (br.s, 1H), 4.84 (m, 1H), 4.23 (m, 1H), 3.75(m, 1H), 3.12-2.5 (m, 10H), 1.81 (m, 4H), 1.43 (s, 9H).

MS (FD) 629 (P+1).

    ______________________________________                                        Analysis:                                                                                 Calculated                                                                           Found                                                      ______________________________________                                        C:            70.79    70.63                                                  H:            7.55     7.31                                                   N:            8.92     9.21                                                   ______________________________________                                    

EXAMPLE 34 1S-(1R*, 4R*,5S*)!-N-(1-(1-methylethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinoxalinylacetamide

The title compound was prepared according to the procedures describedabove in Preparations 1, 2 and 3 and Example 1 using the appropriatereactants.

Yield: 51 mg (8%) as a yellowish foam.

¹ H NMR (300 MHz, CDCl₃) d 9.69 (s, 1H), 8.31 (d, 1H), 8.28-7.0 (m,13H), 6.78 (m, 2H), 6.12 (m, 2H), 4.43 (m, 2H), 3.82 (m, 1H), 1.5 (s,9H), 1.0 (dd, 6H). MS (FD) 597 (P+1).

EXAMPLE 35 1S-(1R*, 4R*,5S*)!-N-(1-(1-methylethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-naphthylacetamide

The title compound was prepared according to the procedures describedabove in Preparation 1, 2 and 3 and Example 1 using the appropriatereactants.

Yield: 39.6 mg (11.1%) of a tan colored foam.

¹ H NMR (300 MHz, CDCl₃) d 8.22 (s, 1H), 7.93-6.7 (m, 15H), 6.1 (d, 1H),6.0 (s, 1H), 4.49 (t, 1H), 4.35 (m, 1H), 3.82 (m, 1H), 3.09-2.77 (m,4H), 2.2 (m, 1H), 1.47 (s, 9H), 0.98 (t, 6H).

MS (FD) 594 (P+1).

EXAMPLE 36 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-3-(1,2,3,4-tetrahydroisoquinolinyl)carboxamide (Diastereomer A)

a. dl-3-carboxy-1,2,3,4-tetrahydroisoquinoline hydrochloride

By following the procedures described in Karo et al., J. Pharm. Soc.Japan, 94, 934 (1974) the title compound was prepared fromL-phenylalanine.

b. dl-2-t-butoxycarbonyl-3-carboxy-1,2,3,4-tetrahydroisoquinoline

By following the procedures described in Bodansky and Bodansky, ThePractice of Peptide Syntheses, Vol. 21, 20 (1984) the title compound wasprepared.

c. Pentafluorophenyl ester ofdl-2-Boc-3-carboxy-1,2,3,4-tetrahydroisoquinoline

The title compund was prepared according to the procedures describedabove in Preparation 2.

d. R-(R*, S*,S*)!-N-t-butyl-2-(3-N-aspariginylamino)-2-hydroxypropyl)benzamide

By following the procedures described in Example 28, steps b through d,the title compound was prepared.

e. dl- 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl-3-(2-t-butoxycarbonyl-1,2,3,4-tetrahydroisoquinolinyl carboxamide

By following the procedures described in Preparation 3, except usingN-methymorpholine rather than sodium bicarbonate and usingdichloroethane rather than dioxane and water, the title compound wasprepared.

f. dl- 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl-3-(1,2,3,4-tetrahydroisoquinolinylcarboxamide

By following the procedures described in Bodansky and Bodansky, thepractice of peptide synthesis, Vol. 21, (1984) for the removal of a Bocgroup, the title compound was prepared.

g. Separation of Diastereomers

The mixture of diastereomers afforded in step f, above, were separatedby chromotography on silica gel using CHCL₃ /CH₃ OH (95:5;V:V) aseluant. Collection of fractions 2-4 afforded a single diastereomerdenoted as Diastereomer A, the configuration of which around the fourthchiral atom has not been assigned.

Yield: 29 mg (38%) of a foam

¹ H NMR (300 MHz, CDCl₃) d 8.25 (d, 1H), 7.4-7.0 (m, 13H), 6.35 (br.s,1H), 6.22 (s, 1H), 5.58 (br.s, 1H), 4.68 (9, 1H), 4.29 (m, 1H), 3.95 (m,2H), 3.75 (m, 1H), 3.52 (m, 1H), 3.1-2.43 (m, 8H), 1.47 (s, 9H).

MS (FD) 614 (P+2).

EXAMPLE 37 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-3-(1,2,3,4-tetrahydroisoquinolinyl)carboxamide (Diastereomer B)

The title compound was prepared along with Example 41 (Diastereomer A)according to steps a through f. Separation of Diastereomer B wasachieved according to Example 41, Step g. Fractions 7-10 were collectedand denoted Diastereomer B, the configuration of which around the fourthchiral atom has not been assigned.

Yield: 32 mg (41%) as a foam.

¹ H NMR (300 MHz, CDCl₃) d 8.16 (d, 1H,) 7.46-6.9 (m, 13H), 6.62 (br.s,1H), 6.37 (br.s, 1H), 5.9 (br.s, 1H), 4.79 (9, 1H), 4.28 (m, 1H),3.92-3.75 (m, 3H), 3.41 (m, 1H, CH), 3.03-2.5 (m, 8H), 1.42 (s, 9H).

MS (FD) 614 (P+2).

EXAMPLE 38 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-(1-methylpropyl)-6-(N-quinolin-2-ylmethoxycarbonylamino)hexyl)benzamide

The title compound was prepared according to the general procedureoutlined in Example 1. Flash chromatography with 3% MeOH/CH₂ Cl₂afforded 0.15 g of the product (68%) as a white solid.

¹ H NMR (300 MHz, CDCl₃) d 0.72 (m, 6H), 1.45 (s, 9H), 1.00-1.80 (m,3H), 2.70-3.10 (m, 6H), 3.79 (m, 1H), 3.99 (m, 1H), 4.35 (m, 1H), 5.35(br.s, 1H), 5.43 (d, J=8 Hz, 1H), 6.24 (br.s, 1H), 6.66 (d, J=9 Hz, 1H),7.05-7.38 (m, 9H), 7.40 (d, J=8 Hz, 1H), 7.49 (t, J=8 Hz, 1H), 7.65 (t,J=8 Hz, 1H), 7.75 (d, J=8 Hz, 1H), 7.96 (d, J=8 Hz, 1H), 8.10 (d, J=8Hz, 1H).

IR (CHCl₃) 3429, 2970, 1725, 1645, 1601, 1515 cm⁻¹.

MS (FD) m/e 639 (M+).

Analytical calc'd for C₃₈ H₄₆ N₄ O₅ : C 71.45, H 7.26, N 8.77; found C71.42, H 7.36, N 8.70.

EXAMPLE 39 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-(1-methylpropyl)-7-aza-8-oxo-8-(N-methyl-N-quinolin-2-ylmethylamino)octyl)benzamide

The title compound was prepared according to the general procedureoutlined in Example 1. Flash chromatography with 3% MeOH/CH₂ Cl₂afforded 40 mg of the product (58% ) as a white solid.

¹ H NMR (300 MHz, CDCl₃) d 0.80 (m, 6H), 1.46 (s, 9H), 1.00-1.97 (m,3H), 2.70-3.10 (m, 4H), 3.02 (s, 3H), 3.75 (m, 1H), 4.13 (t, J=6 Hz,1H), 4.23 (m, 1H), 4.65 (s, 2H), 5.85 (d, J=5 Hz, 1H), 6.22 (s, 1H),6.51 (d, J=9 Hz, 1H), 7.00-7.40 (m, 12H), 7.55 (t, J=8 Hz, 1H), 7.71 (t,J=8 Hz, 1H), 7.80 (d, J=8 Hz, 1H), 8.15 (d, J=8 Hz, 1H).

MS (FD)m/e 652 (M+).

Analytical calc'd for C₃₉ H₄₉ N₅ O₄ : C 71.86, H 7.58, N 10.74; found C71.60, H 7.86, N 10.49.

EXAMPLE 40 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)fur-3-yl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Flash chromatography with 2-10% MeOH/CH₂ Cl₂afforded 80 mg of the product (44%) as a tan solid.

¹ H NMR (300 MHz, CDCl₃) d 1.42 (s, 9H), 2.78 (dd, J=8 Hz, 1H),2.85-3.10 (m, 5H), 3.74 (m, 1H), 4.25 (m, 1H), 4.97 (m, 1H), 5.38 (br.s,1H), 5.91 (br.s, 1H), 6.49 (br.s, 1H), 6.86-7.03 (m, 5H), 7.18-7.32 (m,4H), 7.65 (t, J=8 Hz, 1H), 7.82 (t, J=8 Hz, 1H), 7.91 (d, J=8 Hz, 1H),8.23 (m, 2H), 8.38 (d, J=8 Hz, 1H), 9.25 (d, J=8 Hz, 1H).

IR (CHCl₃) 1681, 1600, 1528, 1500 cm⁻¹.

MS (FAB) mass 600.2839 (M+H).

Analytical calc'd for C₃₃ HB₇ N₅ O₆ : C 66.10, H 6.22, N 11.68; found C65.93, H 6.36, N 11.40.

EXAMPLE 41 1S-(1R*, 4R*. 5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)thien-3-yl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. HPLC with 70% MeOH/1% NH₄ OAc in water afforded90 mg of the product (50%) as a white solid.

¹ H NMR (300 MHz, CDCl₃) d 1.42 (s, 9H), 2.75 (dd, J=7 Hz, 1H),2.95-3.20 (m, 5H), 3.77 (m, 1H), 4.27 (m, 1H), 4.95 (m, 1H), 5.38 (br.s,1H), 5.93 (br.s, 1H), 6.41 (br.s, 1H), 6.95 (m, 5H), 7.20 (m, 4H), 7.66(t, J=8 Hz, 1H), 7.81 (t, J=8 Hz, 1H), 7.90 (d, J=8 Hz, 1H), 8.20 (m,2H), 8.34 (d, J=8 Hz, 1H), 9.25 (d, J=8 Hz, 1H).

IR (KBr) 3302, 1667, 1498, 1427 cm⁻¹.

MS (FAB) mass 616.2591 (M+H), (FD) m/e 615 (M+).

Analytical calc'd for C₃₃ H₃₇ N₅ O₅ S: C 64.37, H 6.06, N 11.37; found C64.60, H 5.98, N 11.42.

EXAMPLE 42 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)benzofur-3-yl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Flash chromatography with 5% MeOH/CH₂ Cl₂afforded 0.25 g of the product (86%) as a white solid.

¹ H NMR (300 MHz, CDCl₃) d 1.42 (s, 9H), 2.75-3.40 (m, 6H), 3.80 (m,1H), 4.37 (m, 1H), 5.00 (m, 1H), 5.61 (br.s, 1H), 6.30 (br.s, 1H), 6.61(br.s, 1H), 6.80-7.40 (m, 11H), 7.62 (t, J=8 Hz, 1H), 7.75 (t, J=8 Hz,1H), 7.85 (d, J=8 Hz, 1H), 8.20 (m, 2H), 8.27 (d, J=8 Hz, 1H), 9.27 (d,J=8 Hz, 1H).

IR (KBr) 3411, 1680, 1603, 1523, 1500 cm⁻¹.

MS (FD)m/e 650 (M+).

Analytical calc'd for C₃₇ H₃₉ N₅ O₆ : C 68.40, H 6.05, N 10.78; found C68.19, H 6.01, N 11.07.

EXAMPLE 43 1S-(1R*, 4R*,5S*)!-N-(1-(2-hydroxy-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The final coupling reaction was accomplished using an activatedpentafluoro ester prepared according to the procedures described inPreparations 2 and 3.

Yield: 40 mg (40%) of a colorless solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.38 (s, 9H), 2.58-3.05 (m, 6H), 3.61(br.s, 1H), 3.84 (m, 1H), 4.79 (m, 1H), 5.88 (s, 1H), 6.92 (t, J=8 Hz,1H), 7.02 (t, J=8 Hz, 2H), 7.12-7.37 (m, 6H), 7.75 (t, J=10 Hz, 2H),7.90 (t, J=10 Hz, 2H), 7.98 (d, J=10 Hz, 2H), 8.07-8.23 (m, 4H), 8.60(d, J=10 Hz, 1H), 8.89 (d, J=10 Hz, 1H), 12.40 (s, 1H).

MS (FD) m/e 613 (25), 612 (95), 611 (100).

Analytical calc'd for C₃₅ H₃₈ N₄ O₆ : C 68.84, H 6.27, N 9.17; found C68.61, H 6.44, N 9.10.

EXAMPLE 44 1S-(1R*, 4R*,5S*)!-N-(1-(3-hydroxy-3-oxopropyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The final coupling reaction was accomplished using an activatedpentafluoro ester prepared according to the procedures described inPreparations 2 and 3.

Yield: 0.58 g (98%) of a colorless solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.34 (s, 9H), 1.96 (m, 2H), 2.17 (t, J=6Hz, 2H), 2.60-2.76 (m, 2H), 2.88 (d, J=15 Hz, 1H), 3.02 (d, J=15 Hz,1H), 3.91 (br.s, 1H), 4.07 (br.s, 1H), 4.48 (m, 1H), 5.83 (br.s, 1H),6.98 (t, J=9 Hz, 1H), 7.10 (t, J=9 Hz, 2H), 7.15-7.35 (m, 6H), 7.72 (t,J=9 Hz, 1H), 7.87 (t, J=9 Hz, 1H), 8.03-8.20 (m, 5H), 8.50 (d, J=9 Hz,1H), 8.72 (d, J=9 Hz, 1H). MS (FD) m/e 626 (15), 625 (35), 220 (8).

Analytical calc'd for C₃₆ H₄₀ N₄ O₆ : C 69.21, H 6.45, N 8.97; found C68.93, H 6.32, N 8.95.

EXAMPLE 45 1S- 1R*, 4R*,5S*)!-N-(1-(2-(N-2-pyridylmethyl)amino)-2-oxoethyl!-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1, using 0.2 g (0.33 mmol) of 1S-(1R*, 4R*,5S*)!-N-(1-(2-hydroxy-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide, from Example 48, and 0.07 mL (0.66 mmol) of2-aminomethylpyridine. The coupling agent for the reaction wascarbonyldiimidazole. Flash chromatography using a silica column with2.5% MeOH/EtOAc containing a trace of NH4OH afforded 33 mg (14%), of thedesired product as a single diastereomer, as determined by 300 MHz ¹ HNMR.

¹ H NMR (300 MHz, CDCl₃) d 1.48 (s, 9H), 2.77-3.16 (m, 6H), 3.80 (br.s,1H), 4.33 (m, 1H), 4.59 (m, 2H), 5.02 (m, 1H), 5.90 (br.s, 1H), 6.23 (s,1H), 6.90-7.09 (m, 4H), 7.15 (m, 1H), 7.20-7.42 (m, 8H), 7.57-7.70 (m,2H), 7.82 (t, J=8 Hz, 1H), 7.91 (d, J=8 Hz, 1H), 8.18-8.40 (m, 4H), 9.35(d, J=8 Hz, 1H).

MS (FD) m/e 701 (100), 700 (15), 481 (28), 220 (90).

Analytical calc'd for C₄₁ H₄₄ N₆ O₅ ·2.2H₂ O: C 66.50, H 6.56, N 11.40;found C 66.35, H 6.20, N 11.00.

EXAMPLE 46 1S-(1R*, 4R*,5S*)!-N-(1-(2-(N-2-imidazolylmethyl)amino)-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1, using 0.50 g (0.82 mmol) of 1S-(1R*, 4R*,5S*)!-N-(1-(2-hydroxy-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide, from Example 48, and 0.28 g (1.64 mmol) ofimidazolemethylamine dihydrochloride. Flash chromatography with 5%MeOH/EtOAc containing a trace of NH₄ OH afforded 0.26 g (46%) of acolorless solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.33 (s, 9H), 2.69 (m, 5H), 2.95 (dd, J=30Hz, 14, 2H), 3.62 (br.s, 1H), 3.88 (br.s, 1H), 4.22 (d, J=10 Hz, 2H),4.80 (br. s, 1H), 6.82 (br. s, 1H), 6.87 (t, J=8 Hz, 1H), 7.05 (t, J=8Hz, 2H), 7.15-7.25 (m, 4H), 7.30 (t, J=7 Hz, 2H), 7.74 (t, J=7 Hz, 1H),7.89 (t, J=9 Hz, 1H), 7.98-8.14 (m, 6H), 8.45 (t, J=6 Hz, 1H), 8.58 (d,J=9 Hz, 1H), 8.96 (d, J=9 Hz, 1H).

MS (FD) m/e 691 (80), 690 (100).

Analytical calc'd for C₃₉ H₄₃ N₇ O₅ : C 67.91, H 6.28, N 14.21; found C67.67, H 6.50, N 13.98.

EXAMPLE 47 1S-(1R*, 4R*,5S*)!-N-(1-(2-(N-benzyloxy)amino)-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(3-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 51. Flash chromatography with 10% MeOH/CH₂ Cl₂afforded 0.189 g (92%) of a white solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.35 (s, 9H), 2.52 (s, 1H), 2.61-2.73 (m,2H), 2.87-3.03 (m, 2H), 3.60-3.64 (m, 1H), 3.85-3.89 (m, 1H), 4.64 (d, j=5.9 Hz, 1H), 4.78 -4.83 (m, 1H), 5.84 (d, J=5.6 Hz, 1H), 6.90-6.95 (m,1H), 7.03 (t, J=7.5 Hz, 2H), 7.13-7.37 (m, 13H), 7.73 (t, J=7.5 Hz, 1H),7.86-7.91 (m, 1H), 8.01-8.20 (m, 5H), 8.60 (d, J=8.6 Hz, 1H), 8.83 (d,J=8.5 Hz, 1H), 11.10 (s, 1H).

IR (KBr) 3319, 2929, 1652, 1522, 1498, 1454, 1366, 1221, 846, 748, 699cm⁻¹.

MS (FD) m/e 717 (M+1, 24), 632 (21), 610 (100), 593 (9), 518 (6), 389(17), 220 (68).

Analytical calc'd for C₄₂ H₄₅ N₅ O₆ : C 70.47, H 6.34, N 9.78; found C70.77, H 6.63, N 9.66.

EXAMPLE 48 1S-(1R*, 4R*,5S*)!-N-(1-(2-(N-pyrid-4-yl)amino)-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 51. Flash chromatography with 3-10% MeOH/CH₂ Cl₂afforded 0.007 g (3%) of a white solid.

¹ H NMR (300 MHz, (DCl₃) d 1.46 (s, 9H), 2.76-3.14 (m, 7H), 3.74-3.83(m, 1H), 4.33-4.41 (m, 1H), 5.03-5.09 (m, 1H), 5.97 (s, 1H), 6.10-6.14(m, 1H), 6.88 (d, J=7 Hz, 1H), 6.94-7.03 (m, 3H), 7.18 (d, J=9 Hz, 2H),7.26 (s, 1H), 7.33-7.43 (m, 4H), 7.66 (t, J=9 Hz, 1H), 7.80 (t, J=9 Hz,1H), 7.92 (d, J=9 Hz), (H), 8.17 (d, J=9 Hz, 1H), 8.28 (s, 1H), 8.32 (s,1H) 8.37 (d, J=8 Hz, 2H), 8.93 (s, 1H), 9.13 (d, J=8 Hz, 1H).

MS (FD) m/e 688 (M+I, 100), 220 (22).

EXAMPLE 49 1S-(1R*, 4R*,5S*)!-N-(1-(2-(N-2-benzimidazolylmethyl)amino)-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 51. Flash chromatography with 3-10% MeOH/CH₂ Cl₂afforded 0.199 g (82%) of a yellow foam.

¹ H NMR (300 MHz, DMSO-d₆) d 1.34 (s, 9H), 2.63-3.30 (m, 5H), 3.60-3.65(m, 1H), 3.88-3.91 (m, 1H), 4.43 (d, J=5.6 Hz, 2H), 4.84 (dd, J=7.8 Hz;6.0, 1H), 5.87 (d, J=5.3 Hz, 1H), 6.92-7.47 (m, 14H), 7.69-7.75 (m, 1H),7.85 (t, J=7.7 Hz, 1H), 8.03-8.20 (m, 6H), 8.57 (d, J=8.6 Hz, 1H), 8.65(m, 1H), 9.00 (d, J=8.2 Hz, 1H).

MS (FD) m/e 743 (M+i), 740 (29), 739 (50), 354 (100).

Analytical calc'd for C₄₃ H₄₅ N₇ O₅ ·0.4H₂ O: C 69.13, H 6.18, N 13.12;found C 68.84, H 6.35, N 13.50.

EXAMPLE 50 1S-(1R*, 4R*,5S*)!-N-(1-(2-(N-(N-3-pyridyl)amino)amino)-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 51. Flash chromatography with 3-10% MeOH/CH₂ Cl₂afforded 0.171 g (74.5%) of an orange solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.35 (s, 9H), 2.63-2.82 (m, 4H), 2.88-3.02(m, 2H), 3.59-3.63 (m, 1H), 3.87-3.90 (m, 1H), 4.80-4.88 (m, 1H), 5.82(d, J=5.4 Hz, 1H), 6.46-6.51 (m, 2H), 6.93-6.97 (m, 1H), 7.05 (t, J=7.4Hz, 1H), 7.16-7.22 (m, 4H), 7.28-7.32 (m, 2H), 7.71-7.75 (m, 1H),7.86-7.91 (m, 2H), 8.03 (d, J=8.9 Hz, 1H), 8.08-8.19 (m, 5H), 8.61 (d,J=8.5 Hz, 1H), 8.92 (d, J=8.4 Hz, 1H), 9.79 (s, 1H).

MS (FD) m/e 702 (M--, 100).

Analytical calc'd for C₄₀ H₄₃ N₇ OS: C 68.46, H 6.18, N 13.97; found C65.74, H 6.26, N 14.22.

EXAMPLE 51 1S-(1R*, 4R*.5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-cyclohexylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Flash chromotography with 5% MeOH in CH₂ Cl₂containing a trace of NH₄ OH afforded 0.63 g (71%) of an off-whitesolid.

¹ H NMR (300 MHz, CDCl₃) d 1.44 (s, 9H), 0.71-1.66 (m, 8H), 1.82 (d,J=11 Hz, 1H), 5.60 (s, 1H), 6.33 (br.s), 2H), 7.11 (d, J=10 Hz, 1H),7.18-7.41 (m, 6H), 7.66 (t, J=9 Hz, 1H), 7.81 (t, J=9 Hz, 1H), 7.90 (d,J=9 Hz, 1H), 8.27 (t, J=9 Hz, 2H), 8.37 (d, J=9 Hz, 1H), 9.48 (d, J=9Hz, 1H).

MS (FD) m/e 616 (100), 396 (15), 395 (33), 394 (8), 221 (18), 220 (100),192 (20), 191 (88).

Analytical calc'd for C₃₅ H₄₅ N₅ O₅ : C 68.27, H 7.37, N 11.37; found: C68.47, H 7.48, N 11.62.

EXAMPLE 52 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-8-fluoroquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1, using carbonyldiimidazole as a coupling agent.Yield: 158 mg (25%).

¹ H NMR (300 MHz, CDCl₃) d 1.4 (s, 9H), 2.7-3.1 (m, 6H), 3.7-3.8 (m,1H), 4.3-4.4 (m, 1H), 4.9-5.0 (m, 1H), 5.44 (m, 1H), 6.0-6.1 (m, 2H),6.9-7.7 (m, 14H), 8.3-8.4 (2d, 2H), 9.2 (d, 1H).

MS (FD) 627.

Analytical calc'd for C₃₅ H₃₈ N₅ O₅ F: C 66.97, H 6.10, N 11.16; found C67.21, H 6.22, N 11.03.

EXAMPLE 53 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-4-chloroquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 57. Yield: 29.2 mg (9%).

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H), 2.7-3.1 (m, 6H), 3.7-3.8 (m,1H), 4.3-4.4 (m, 1H), 4.9-5.0 (m, 1H), 5.5 (m, 1H), 6.0 (m, 2H), 6.9-8.3(m, 17H), 9.2 (d, 1H).

MS (FD) 644.

Analytical calc'd for C₃₅ H₃₈ N₅ O₅ Cl: C 65.26, H 5.95, N 10.87; foundC 65.55, H 6.04, N 10.85.

EXAMPLE 54 1S-(1R*, 4R*,5S*)!-N-(1-(3-amino-3-oxopropyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Yield: 15 mg (12.6%).

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H), 2-2.3 (m, 4H), 2.8-3.1 (m, 4H),3.91 (m, 1H), 4.39 (m, 1H), 4.64 (m, 1H), 5.6 (br.s, 1H), 6.03 (br.d,1H), 6.2 (br.s, 1H), 6.5 (br.s, 1H), 6.97 (m, 1H), 7.08-7.4 (m, 9H),7.65 (t, 1H), 7.8 (t, 1H), 7.89 (d, 1H), 8.14-8.32 (m, 3H), 8.8 (d, 1H).

MS (FD) 624 (P+2), 603, 532, 403, 220.

Analytical calc'd for C₃₆ H₄₁ N₅ O₅ : C 71.36, H 6.80, N 9.00; found C70.61, H 6.92, N 8.82.

EXAMPLE 55 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-6-chloroquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 57. Yield: 40 mg (6%).

¹ H NMR (300 MHz, CDCl₃) d 1.4 (s, 9H), 2.6-3.1 (m, 6H), 3.7-3.8 (m,1H), 4.3-4.4 (m, 1H), 4.9-5.0 (m, 1H), 5.4 (m, 1H), 5.9-6.0 (m, 2H),6.9-8.25 (m, 16H), 9.2 (d, 1H).

MS (FAB) 644.

Analytical calc'd for C₃₅ H₃₈ N₅ O₅ Cl: C 65.26, H 5.95, N 10.87; foundC 66.38, H 6.11, N 10.86.

EXAMPLE 56 1S-(1R*, 4R*, 5S*)!-N-(1-(3-amino-3-oxopropyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-naphthylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Yield: 45 mg (35%).

¹ H NMR (300 MHz, CDCl₃) d 1.49 (s, 9H), 2.1-2.3 (m, 4H), 2.8-3.13 (m,4H), 3.87 (m, 1H), 4.35 (m, 1H), 4.66 (9, 1H), 5.82 (br.s, 1H), 6.04 (d,1H), 6.29 (s, 1H), 6.57 (br.s, 1H), 6.98-7.93 (m, 17H), 8.29 (s, 1H).

MS (FD) 623 (P+), 605, 531, 220.

Analytical calc'd for C₃₇ H₄₂ N₄ O₅ : C 71.36, H 6.80, N 9.00; found C70.61, H 6.92, N 8.82.

EXAMPLE 57 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-(2-amino-2-oxoethyl)-7-aza-8-oxo-8-(N-benzylamino)octyl)benzamide

A solution of 60 mg (0.131 mmol) of 2R-(2R*, 3S*, 6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-(2-amino-2-oxoethyl)-6-aminohexyl)benzamideand 0.02 mL (0.158 mmol) of benzylisocyanate in 2 mL of THF wasprepared, under nitrogen. This solution was stirred at room temperaturefor 2 hours, resulting in the formation of a colorless precipitate Thesolid was collected by filtration, washed with cold THF and dried invacuo to afford 15 mg of a colorless solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.32 (s, 9H), 2.29-3.0 (m, 6H), 3.57 (m,1H), 3.8 (m, 2H), 4.18 (d, 1H), 4.32 (m, 1H), 5.79 (d, 1H), 6.14 (d,1H), 6.62 (t, 1H), 6.82 (br.s, 1H), 7.03-7.3 (m, 14H), 7.71 (d, 1H),8.19 (s, 1H).

MS (FD) 588 (P), 481, 367, 220.

Analytical calc'd for C₃₃ H₄₁ N₅ O₅ : C 67.44, H 7.03, N 11.92; found C67.20, H 7.13, N 12.18.

EXAMPLE 58 2R-(2R*, 3S*,6S*)!-N-t-butyl-2-(2-hydroxy-3-phenylmethyl-4-aza-5-oxo-6-(2-amino-2-oxoethyl)-7-aza-8-oxo-8-(N-naphth-1-ylmethylamino)octyl)benzamide

The title compound was prepared according to the general procedureoutlined in Example 62 to afford 116.5 mg (69%) of a colorless solid.

¹ H NMR (300 MHz, DMSO-d₆) d 1.39 (s, 9H), 2.3-3.0 (m, 6H), 3.55 (m,1H), 3.84 (m, 1H), 4.35 (9,1H), 4.65 (d, 2H), 5.82 (d, 1H), 6.19 (d,1H), 6.7 (t, 1H), 6.86 (br.s, 1H), 7.41-7.4 (m, 11H), 7.54 (m, 2H), 7.73(d, 1H), 7.8 (m, 1H), 7.91 (m, 1), 8.04 (m, 1H), 8.2 (s, 1H).

MS (FD) 638 (P+1).

Analytical calc'd for C₃₇ H₄₃ N₅ O₅ : C 69.68, H 6.80, N 10.98; found C69.54, H 6.70, N 10.74.

EXAMPLE 59 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-6,8-difluoroquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 57. Yield: 51.3 mg (8%).

¹ H NMR (300 MHz, CDCl₃) d 1.4 (s, 9H), 2.6-3.1 (m, 6H), 3.7-3.8 (m,1H), 4.3-4.4 (m, 1H), 4.9-5.0 (m, 1H), 5.3-5.4 (m, 1H), 5.9 (m, 1H), 6.0(m, 1H), 6.6-7.4 (m, 13H), 8.3 (s, 2H), 9.2 (d, 1H).

MS (FD) 646 (M+1).

Analytical calc'd for C₃₅ H₃₇ N₅ OSF₂ : C 65.10, H 5.78, N 10.84; foundC 65.64, H 6.47, N 11.68.

EXAMPLE 60 1S-1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-4-chloro-8-isopropylquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 57. Yield: 38.5 mg (3%).

¹ H NMR (300 MHz, CDCl₃) d 1.4-1.5 (m, 15H), 2.6-3.1 (m, 6H), 3.7-3.8(m, 1H), 4.3-4.4 (m, 2H), 4.9-5.0 (m, 1H), 5.3 (m, 1H), 5.9-6.0 (m, 2H),6.9-7.4 (m, 10H), 7.7-7.8 (m, 2H), 8.1-8.2 (d, 2H), 8.3 (s, 1H), 9.2 (d,1H).

MS (FD) 686.

EXAMPLE 61 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-8-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 57.

¹ H NMR (300 MHz, DMSO-d₆) d 1.3 (s, 9H), 2.2-3.0 (m, 6H), 3.6 (m, 1H),3.8 (m, 1H), 4.3-4.4 (m, 1H), 5.8 (d, 1H), 6.4 (d, 1H), 6.8 (s, 1H),7.1-7.4 (m, 16H), 7.6 (d, 1H), 8.2 (d, 1H).

MS (FD) 644.

EXAMPLE 62 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-8-chloroquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 57. yield: 62.5 mg (18%).

¹ H NMR (300 MHz, CDCl₃) d 1.4 (s, 9H), 2.2-3.1 (m, 6H), 3.7 (m, 1H),4.3-4.4 (m, 1H), 4.8-4.9 (m, 1H), 5.4 (m, 1H), 5.9 (m, 1H), 6.0 (m, 1H),6.9-8.4 (m, 14H), 9.4 (d, 1H).

MS (FD) 644.

Analytical calc'd for C₃₅ H₃₈ N₅ O₅ Cl: C 65.26, H 5.95, N 10.87; foundC 65.50, H 6.04, N 10.94.

EXAMPLE 63 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-5,7-dimethylquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Chromatography over a silica column with 0-5%MeOH in CHCl₃ afforded 0.52 g (74%).

¹ H NMR (300 MHz, CDCl₃) d 1.48 (s, 9H), 2.57 (s, 3H), 2.72 (s, 3H),2.71-3.18 (m, 5H), 3.68-3.8 (m, 2H), 4.25-4.39 (m, 1H), 4.91-5.0 (m,1H), 5.36 (s, 1H), 6.07 (s, 2H), 6.9-7.13 (m, 5H), 7.20-7.42 (m, 8H),7.92 (s, 1H), 8.21 (d, J=8 Hz, 1H), 8.48 (d, J=8 Hz, 1H), 9.38 (d, J=5Hz, 1H). MS (FD) 638, 546.

Analytical calc'd for C₃₇ H₄₃ N₅ O₅ : C 69.68, H 6.80, N 10.98; found C69.83, H 6.99, N 10.93.

EXAMPLE 64 1S-(1R*, 4R*, 5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-4-methoxyquinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Chromatography over a silica column with 0-5%MeOH in CHCl₃ afforded 560 mg (80%).

¹ H NMR (300 MHz, CDCl₃) d 1.47 (s, 9H), 2.68-3.13 (m, 6H), 3.72-3.82(m, 1H), 4.16 (s, 3H), 4.23-4.39 (m, 1H), 4.92-5.02 (m, 1H), 5.48 (br.s,1H), 5.85-6.24 (m, 3H), 6.92-7.41 (m, 9H), 7.58-7.84 (m, 3H), 9.32-9.48(br.s, 1H).

MS (FD) 640, 420, 220.

EXAMPLE 65 1S-(1R*, 4R*,5S*)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-5-methylphenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Chromatography over a silica column with 1-5%MeOH in CH₂ Cl₂ afforded a white solid.

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H), 2.35 (s, 3H), 2.68-3.12 (m,6H), 3.70-3.78 (m, 1H), 4.25-4.38 (m, 1H), 4.95-5.02 (m, 1H), 6.00 (s,1H), 6.05 (br.s, 1H), 6.89-7.30 (m, 8H), 7.64-7.96 (m, 3H), 8.2-8.4 (m,3H), 9.28 (d, J=6 Hz, 1H).

MS (FAB) 624, 607, 355.

Analytical calc'd for C₃₆ H₄₁ N₅ O₅ : C 69.32, H 6.62, N 11.23; found C69.50, H 6.71, N 11.31.

EXAMPLE 66 1S-(1R*, 4R*,5S*)!-N-(1-(3-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-4-(2,2-dimethylethyl)quinolin-2-ylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1. Chromatography over a silica column with 0-5%MeOH in CHCl₃ afforded 641 mg (87%).

¹ H NMR (300 MHz, CDCl₃) d 0.88 (t, J=7 Hz, 3H), 1.45 (s, 9H), 1.73 (d,J=7 Hz, 3H), 1.69-1.79 (m, 2H), 2.68-3.12 (m, 7H), 3.70-3.79 (m, 1H),4.26-4.37 (m, 1H), 4.92-5.01 (m, 1H), 5.55 (br.s, 1H), 6.12-6.22 (m,2H), 6.91-7.40 (m, 10H), 7.63-7.72 (m, 2H), 8.13-8.31 (m, 3H), 9.23 (d,J=8 Hz, 1H).

MS (FD) 666, 573,445.

Analytical calc'd for C₂₉ H₄₇ N₅ O₅ : C 70.35, H 7.12, N 10.52; found C70.09, H 7.03, N 10.44.

EXAMPLE 67 1S-(1R*, 4R*,5S*)!-N-(1-(4-imidazolylmethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-4-methylphenyl)-4-hexyl)-2-quinolinylcarboxamide

The final coupling reaction was accomplished using an activatedpentafluoro ester prepared according to the procedures described inPreparations 2 and 3.

Yield: 72 mg (31%) of a colorless solid.

¹ H NMR (300 MHz, CDCl₃) d 1.45 (s, 9H), 2.26 (s, 3H), 2.73-2.80 (m,2H), 2.94-2.99 (m, 2H), 3.13-3.23 (m, 2H), 3.90-3.95 (m, 1H), 4.29-4.40(m, 1H), 4.89-4.97 (m, 1H), 6.68 (s, 1H), 6.79 (s, 1H), 6.85-7.15 (m,10H), 7.49-7.60 (m, 3H), 7.72 (t, J=7.5 Hz, 1H), 7.79 (d, J=8 Hz, 1H),8.03-8.14 (m, 2H), 8.18 (d, J=8 Hz, 1H), 8.85 (d, J=6.3 Hz, 1H).

EXAMPLE 68 1S-(1R*, 4R*,5S,)!-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)-5-isopropylphenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1.

Yield: 85%.

¹ H NMR (300 MHz, CDCl₃) d 1.20-1.30 (m, 6H), 1.45 (s, 9H), 2.60-3.20(s, 7H), 3.70-3.80 (m, 1H), 4.25-4.40 (m, 1H), 4.90-5.00 (m, 1H),5.50-6.20 (m, 4H), 6.80-7.30 (m, 9H), 7.55-8.30 (m, 6H), 9.15-3.05 (m,1H).

MS (FD) 652 (M+100).

Analytical calc'd for C₃₈ H₄₅ N₅ O₅ : C 70.02, H 6.96, N 10.74; found C69.83, H 6.98, N 10.62.

EXAMPLE 69 1S-(1R*, 4R*, 5S*) !-N-(1-(2-amino-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-(1-methylcyclopent-1-yl)amino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 1.

The title compound was prepared according to the general procedureoutlined in Example 1.

Yield: 36%

¹ H NMR (300 MHz, DMSO-d₆) d 1.22 (s, 3H), 1.25-1.80 (m, 6H), 2.04-2.21(m, 2H), 2.60-2.80 (m, 4H), 2.90-3.10 (m, 2H), 3.60-3.70 (m, 1H),3.80-3.95 (m, 1H), 4.70-4.80 (m, 1H), 5.85 (d, 1H), 6.9-9.00 (m, 19H).

MS (FD) 636 (M+100).

Analytical calc'd for C₃₇ H₄₁ N₅ O₅ : C 69.90, H 6.50, N 11.02; found C69.97, H 6.64, N 10.95.

EXAMPLE 70 1S-(1R*, 4R*,5*)!-N-(1-(2-(N-indol-3-ylmethyl)amino)-2-oxoethyl)-2-oxo-3-aza-4-phenylmethyl-5-hydroxy-6-(2-(1-t-butylamino-1-oxomethyl)phenyl)hexyl)-2-quinolinylcarboxamide

The title compound was prepared according to the general procedureoutlined in Example 51. Flash chromatography with 3-10% MeOH/CH₂ Cl₂,followed by preparatory HPLC with 35% H₂ O in MeOH afforded 0.087 g(48%) of a light yellow foam.

¹ H NMR (300 MHz, (DMSO-d₆) d 1.35 (s, 9H), 2.61-2.74 (m, 4H), 2.92-3.04(m, 2H), 3.60-3.65 (m, 1H), 3.86-3.89 (m, 1H), 4.30-4.34(m, 1H),4.30-4.34 (m, 2H), 4.77-4.82 (m, 1H), 5.85 (d, J=5.5 Hz, 1H), 6.69 (t,J=7.4 Hz, 1H), 6.91-6.97 (m, 2H), 7.06 (t, J=7.4 Hz, 2H), 7.15-7.32 (m,8H), 7.39 (d, J=7.9 Hz, 1H), 7.73 (t, J=7.8 Hz, 1H), 7.87 (t, j =7.8 Hz,1H), 8.04 (d, J=9 Hz., 1H), 8.10 (d, J=8.5 Hz, 2H), 8.17 (d, J=8.5 Hz,1H), 8.21 (br.s, 2H) 8.59 (d, J=8.5 Hz, 1H), 8.89 (d, J=8.1 Hz, 1H),10.82 (s, 1H).

MS (FD) m/e 739 (M+1, 35), 738 (100). Analytical calc'd for C₄₄ H₄₆ N₆O₅ ·0.50H₂ O: C 70.66, H 6.33, N 11.24; found C 70.28, H 6.32, N 10.98.

By substantially following the procedures described above one skilled inthe art can prepare the compounds of Formula I.

As noted above, the compounds of the present invention are useful forinhibiting HIV protease which is associated with viral componentproduction and assembly. A further embodiment of the present inventionis a method of treating HIV infection comprising administering to amammal in need of treatment an HIV inhibiting dose of a compoundaccording to claim I or a pharmaceutically acceptable salt or solvatethereof. Another embodiment of the present invention is a method forinhibiting HIV replication by administering to a mammal infected withHIV, an HIV protease inhibiting dose (effective amount) of a compoundaccording to Formula I or a pharmaceutically acceptable salt or solvatethereof.

The term "effective amount" as used herein, means an amount of acompound of the present invention which is capable of inhibiting the HIVprotease mediated viral component production and assembly. The HIVprotease inhibition contemplated by the present method includes bothmedical therapeutic and/or prophylactic treatment, as appropriate. Thespecific dose of compound administered according to this invention toobtain therapeutic and/or prophylactic effects will, of course, bedetermined by the particular circumstances surrounding the case,including, for example, the compound administered, the route ofadministration, and the condition being treated. A typical daily dosewill contain a nontoxic dosage level of from about 0.01 mg/kg to about50 mg/kg of body weight of an active compound of this invention.Preferred daily doses generally will be from about 0.05 mg/kg to about20 mg/kg and ideally from about 0.1 mg/kg to about 10 mg/kg.

The compounds can be administered by a variety of routes including oral,rectal, transdermal, subcutaneous, intravenous, intramuscular, andintranasal. The compounds of the present invention are preferablyformulated prior to administration. Therefore, another embodiment of thepresent invention is a pharmaceutical formulation comprising aneffective amount of a compound of Formula I or a pharmaceuticallyacceptable salt thereof and a pharmaceutically acceptable carrier,diluent or excipient therefor.

The active ingredient in such formulations comprises from 0.1% to 99.9%by weight of the formulation. By "pharmaceutically acceptable" it ismeant the carrier, diluent or excipient must be compatible with theother ingredients of the formulation and not deleterious to therecipient thereof.

The present pharmaceutical formulations are prepared by known proceduresusing well known and readily available ingredients. In making thecompositions of the present invention, the active ingredient willusually be admixed with a carrier, or diluted by a carrier, or enclosedwithin a carrier which may be in the form of a capsule, sachet, paper orother container. When the carrier serves as a diluent, it may be asolid, semi-solid or liquid material which acts as a vehicle, excipientor medium for the active ingredient. Thus, the compositions can be inthe form of tablets, pills, powders, lozenges, sachets, cachets,elixirs, suspensions, emulsions, solutions, syrups, aerosols, (as asolid or in a liquid medium), ointments containing, for example, up to10% by weight of the active compound, soft and hard gelatin capsules,suppositories, sterile injectable solutions, sterile packaged powders,and the like.

The following formulation examples are illustrative only and are notintended to limit the scope of the invention in any way. "Activeingredient," of course, means a compound according to Formula I or apharmaceutically acceptable salt thereof.

Formulation 1

Hard gelatin capsules are prepared using the following ingredients:

    ______________________________________                                                      Quantity                                                                      (mg/capsule)                                                    ______________________________________                                        Active ingredient                                                                             250                                                           Starch, dried   200                                                           Magnesium stearate                                                                            10                                                            Total           460          mg                                               ______________________________________                                    

Formulation 2

A tablet is prepared using the ingredients below:

    ______________________________________                                                        Quantity                                                                      (mg/capsule)                                                  ______________________________________                                        Active ingredient 250                                                         Cellulose, microcrystalline                                                                     400                                                         Silicon dioxide, fumed                                                                          10                                                          Stearic acid      5                                                           Total             665         mg                                              ______________________________________                                    

The components are blended and compressed to form tablets each weighing665 mg

Formulation 3

An aerosol solution is prepared containing the following components:

    ______________________________________                                                            Weight                                                    ______________________________________                                        Active ingredient     0.25                                                    Ethanol               25.75                                                   Propellant 22 (Chlorodifluoromethane)                                                               70.00                                                   Total                 100.00                                                  ______________________________________                                    

The active compound is mixed with ethanol and the mixture added to aportion of the propellant 22, cooled to -30° C. and transferred to afilling device. The required amount is then fed to a stainless steelcontainer and diluted with the remainder of the propellant. The valveunits are then fitted to the container.

Formulation 4

Tablets, each containing 60 mg of active ingredient, are made asfollows:

    ______________________________________                                        Active ingredient        60      mg                                           Starch                   45      mg                                           Microcrystalline cellulose                                                                             35      mg                                           Polyvinylpyrrolidone (as 10% solution in water)                                                        4       mg                                           Sodium carboxymethyl starch                                                                            4.5     mg                                           Magnesium stearate       0.5     mg                                           Talc                     1       mg                                           Total                    150     mg                                           ______________________________________                                    

The active ingredient, starch and cellulose are passed through a No. 45mesh U.S. sieve and mixed thoroughly. The aqueous solution containingpolyvinyl-pyrrolidone is mixed with the resultant powder, and themixture then is passed through a No. 14 mesh U.S. sieve. The granules soproduced are dried at 50° C. and passed through a No. 18 mesh U.S.Sieve. The sodium carboxymethyl starch, magnesium stearate and talc,previously passed through a No. 60 mesh U.S. sieve, are then added tothe granules which, after mixing, are compressed on a tablet machine toyield tablets each weighing 150 mg.

Formulation 5

Capsules, each containing 80 mg of active ingredient, are made asfollows:

    ______________________________________                                        Active ingredient  80         mg                                              Starch             59         mg                                              Microcrystalline cellulose                                                                       59         mg                                              Magnesium stearate 2          mg                                              Total              200        mg                                              ______________________________________                                    

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 45 mesh U.S. sieve, and filled into hardgelatin capsules in 200 mg quantities.

Formulation 6

Suppositories, each containing 225 mg of active ingredient, are made asfollows:

    ______________________________________                                        Active ingredient  225         mg                                             Saturated fatty acid glycerides                                                                  2,000       mg                                             Total              2,225       mg                                             ______________________________________                                    

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2 g capacity and allowed to cool.

Formulation 7

Suspensions, each containing 50 mg of active ingredient per 5 ml dose,are made as follows:

    ______________________________________                                        Active ingredient   50         mg                                             Sodium carboxymethyl cellulose                                                                    50         mg                                             Syrup               1.25       ml                                             Benzoic acid solution                                                                             0.10       ml                                             Flavor              q.v.                                                      Color               q.v.                                                      Purified water to total                                                                           5          ml                                             ______________________________________                                    

The active ingredient is passed through a No. 45 mesh U.S. sieve andmixed with the sodium carboxymethyl cellulose and syrup to form a smoothpaste. The benzoic acid solution, flavor and color are diluted with aportion of the water and added, with stirring. Sufficient water is thenadded to produce the required volume.

Formulation 8

An intravenous formulation may be prepared as follows:

    ______________________________________                                        Active ingredient                                                                             100          mg                                               Isotonic saline 1,000        ml                                               ______________________________________                                    

The solution of the above ingredients generally is administeredintravenously to a subject at a rate of 1 ml per minute.

The following experiments were carried out to demonstrate the ability ofthe compounds of the present invention to inhibit HIV protease.

The following abbreviations have the following meanings.

BSA--bovine serum albumin

BOC--t-butyloxycarbonyl

BrZ--2-bromobenzyloxycarbonyl

2-Clz--2-chlorobenzyloxycarbonyl

DCC--dicyclohexylcarbodiimide

DIEA--diisopropylethylamine

DMSO--dimethylsulfoxide

DTT--dithiothreitol

EDTA--ethylenediaminetetraacetic acid

FITC--fluorescein isothiocarbamyl

HEPES--4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

MES--4 morpholineethanesulfonic acid

PAM--phenylacetimidomethyl

TAPS--3- tris(hydroxymethyl)methyl!amino-1-sulfonic acid

TRIS--tris(hydroxymethyl)aminomethane

TOS--p-toluenesulfonyl (tosyl)

Preparation of Substrate

N.sup.α -Biotin-Gly-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gly-Lys(Ne-FITC)-OH

A. Preparation of N^(a) -Biotin-Gly-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Lys-OH

The protected peptide-resin N.sup.α-Boc-Gly-Ser-Gln-Asn-Tyr(BrZ)-Pro-Ile-Val-Gly-Lys(2-C₁ Z)-OCH₂-PAM-resin was synthesized on an Advanced Chemtech Model 200 peptidesynthesizer at 1.5 mmol scale using the standard double-couple protocol.The amino terminal Boc group was removed with 50% CF₃ COOH/CH₂ Cl₂ andthe resuling resin neutralized with 5% diisopropylethylamine (DIEA) inCH₂ Cl₂. Then 1.1 grams of biotin (4.5 mmol) was dissolved in 20 mL ofdimethyl sulfoxide and the solution added to the peptide resin. Then,4.5 mmol of DCC in 9 mL of CH₂ Cl₂ was added to the resin and thereaction mixture brought to 40 mL total volume with 11 mL CH₂ Cl₂. Thecoupling reaction was allowed to run for a total of 5 hours. Thereaction solution was removed, the resin washed with DMSO, DMF and CH₂Cl₂, the resin neutralized with 5% DIEA in CH₂ Cl₂, and the reactionrepeated twice more with the reaction time being extended to 12 hoursper reaction. Ninhydrin analysis of the resin indicated completereaction of the biotin with the glycine amine group. The final peptideresin was washed extensively with DMF and CH₂ Cl₂ and dried. Final yield=4.3 g (98% of theoretical).

B. Deprotection

The peptide was deprotected and cleaved from the resin using 50 ml ofHF/m-cresol (9:1), 0° C., 1 hour. After removal of the HF by vacuumdistillation, the m-cresol was extracted from the reaction mixture with100 mL diethyl ether. The peptide was solubilized in 50% aqueous aceticacid, frozen and lyophilized. Final yield =2.14 g.

C. Purification

The crude N.sup.α -Biotin-Gly-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gly-Lys-OH wasdissolved in 200 mL of 0.1% CF₃ COOH in 95:5 H₂ O:CH₃ CN, filteredthrough a 0.22 micron filter and applied to a 2.2×25 cm. reverse phasecolumn of octadecyl-silica (vydac C-18) which had been equilibrated withthe same buffer. The peptide was eluted with a 855 minute lineargradient of 7.5 to 25% CH₃ CN at 2 mL/minute with collection offractions. Analytical high-performance liquid chromatography wasperformed on a 4.6×250 mm Vydac C-18 column using similar bufferconditions to analyze the column fractions. Column fractions containingthe desired material were pooled, frozen and lyophilized. Finalyield=1.206 g (62% of theory).

Amino acid analysis of the isolated N.sup.α-Biotin-Gly-Ser-Gln-Asn-Tyr-Pro-lle-Val-Gly-Lys-OH gave the followingratios: Asn 1.1; Ser 0.96; Gln 1.1; Pro 1.1; Gly 2.1; Val 0.80; Ile0.78; Tyr 1.1; Lys 1.1; in agreement with theory. Fast-atom bombardmentmass spectrometry gave a molecular ion mass peak of 1288, in agreementwith theory.

D. Labeling

The purified peptide was labeled as follows with a fluorescent marker atthe C-terminal end for use in the assay. N.sup.α-Biotin-Gly-Ser-Gln-Asn-Tyr-Pro-lle-Val-Gly-Lys-OH (1.206 g, 0.936mmoles) was dissolved in 100 mL of 0.1M sodium borate, pH 9.5 withstirring. Then, 3 g of fluorescein isothiocyanate (7.7 mmol) wasdissolved in 15 mL dimethyl sulfoxide and the solution added to thereaction in 10 equal portions over the course of a two hour period. Thereaction was allowed to proceed for a further one hour after the finaladdition had been made. The pH of the solution was adjusted to 3 with 5NHCl. A precipitate formed which was removed from the reaction bycentrifugation.

The pH of the remaining peptide solution was raised to 7.8 with 5N NaOHand the volume adjusted to 200 mL with 0.1M ammonium acetate, pH 7.5.The peptide solution was filtered through a 0.22 micron filter andloaded onto a 2.2×25 cm column of Vydac C-18 which had been equilibratedwith 95:5 0.1M ammonium acetate, pH 7.5:CH₃ CN. The peptide was elutedfrom the column with a 855 minute linear gradient of 5 to 25% CH₃ CN, 2mL/minute with collection of fractions. Analytical HPLC was used toidentify fractions with the desired material, which were then pooled,frozen and lyophilized. Final yield =190.2 mg (12% of theoretical).

Amino acid analysis of the purified peptide gave the following: Asn 1.1;Ser 1.0; Gln 1.1: Pro 1.1; Gly 2.1; Val 0.8; Ile 0.8; Tyr 1.1; Lys 1.0;in agreement with theory. Fast-atom bombardment mass spectrometry gave amolecular ion mass peak of 1678, in agreement with theory.

The following describes the compositions of the reagents (buffers andsolutions) used and referred to in the following evaluations.

    ______________________________________                                        MES-ALB Buffer: 0.05M 4-MES, pH 5.5                                                           0.02M NaCl                                                                    0.002M EDTA                                                                   0.001M DTT                                                                    1.0 mg/ml BSA                                                 TBSA Buffer:    0.02M TRIS                                                                    0.15M NaCl                                                                    1.0 mg/ml BSA                                                 Avidin Coated Beads Solution:                                                                 0.1% solution of Fluoricon Avidin                                             Assay Particles (Avidin conjugated to                                         solid polystyrene beads, 0.6-0.8                                              microns in diameter in TBSA Buffer                            Enzyme Solution:                                                                              27 IU/ml of purified HIV-1 protease in                                        MES-ALB buffer (1 IU equals the                                               amount of enzyme required to hydro-                                           lyze 1 μmol of substrate per                                               minute at 37° C.)                                      ______________________________________                                    

Fluorescence HIV-1 Protease Inhibitor Assay Procedure.

To each well of a round bottom, 96-well plate is added 20 μl of theEnzyme Solution followed by 10 μl of the compound to be evaluated in 20%aqueous DMSO. Purified HIV-1 protease was obtained as described below.The solution is incubated for one hour at room temperature and then 20μl of a solution of the substrate N.sup.α-Biotin-Gly-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gly-Lys(N^(e) -FITC)-OH, inMES-ALB buffer (1.5 μl/ml) is added to each well. The solutions areincubated for 16 hours at room temperature and thereafter to each wellis added 150 μl of MES-ALB buffer.

To each well of a 96-well Pandex plate is added 25 μl of the AvidinCoated Beads Solution. Next, 25 μl of the diluted incubation solutions,described above, are added to each well of the Pandex plate. Thesolutions are mixed well and the plates are loaded into a Pandex®machine, washed, evacuated and read. Sample detection was performed byexcitation at 485 nm and reading the resulting epifluorescence at 535nm.

Inhibition of HIV-1 Protease Produced in E. coli.

The compounds of the present invention were also tested for theirability to block HIV protease processing of Pr48^(gag) (a recombinantpolyprotein containing p 17, p24 and p7).

A. Culture of E. coli K12 L507/DHP10D

Lyophils of E. coli K12 L507/pHP10D are obtained from the NorthernRegional Research Laboratory, Peoria, Ill. 61604, under the accessionnumber NRRL B-18560 (deposited Nov. 14, 1989). The lyophils are decantedinto tubes containing 10 ml LB medium (10 g Bacto-tryptone, 5 gBacto-yeast extract, and 10 g NaCl per liter; pH is adjusted to 7.5) andincubated at 32° C., overnight.

A small portion of the overnight culture was placed on LB-agar (LBmedium with 15 g/l Bacto-agar) plates containing 12.5 μg/ml tetracyclinein a manner so as to obtain a single colony isolate of E. coli K12L507/pHP10D. The single colony obtained was inoculated into 10 ml of LBmedium containing 12.5 μg/ml tetracycline and incubated overnight at 32°C. with vigorous shaking. The 10 ml overnight culture was inoculatedinto LB medium containing 12.5 μg/ml tetracycline and incubated at 32°C. with vigorous shaking until the culture reached mid-log phase.

B. Culture of E. coli K12 L507/pHGAG

Lyophils of E. coli K12 L507/pHGAG are obtained from the NRRL under theaccession number NRRL B-18561 (deposited Nov. 14, 1989). A purifiedcolony of E. coli K 12 L507/pHGAG was isolated, and used as an inoculumfor a culture which was grown to mid-log phase in substantial accordancewith the teaching of Step A above for E. Coli K12 L507/pHP10D.

C. Preparation of Protease and gag fractions

A culture of E. coli K12 L507/pHP10D was grown to mid-log phase at 32°C. in LB media containing 12.5 μg/ml tetracycline. The cultivationtemperature was quickly elevated to 40° C. to induce gene expression,and the cells were allowed to grow for 2.5 hours at this temperaturebefore the culture was quickly chilled on ice. The cells werecentrifuged and the cell pellet was resuspended in 20 ml 50 mmol MESbuffer (pH 6.0) containing 1 mmol EDTA, 1 mmol DTT, 1 mmol PMSF, and 10%glycerol (Buffer A). Cells were lysed by sonication using a FischerModel 300 Dismembrator and a microtip probe. Following centrifugation at27,000 x g, the supernatant was brought to 60 ml with Buffer A andloaded at 1 ml/min onto a QAE-Sepharose column (2.0×19 cm) at 4° C.equilibrated in Buffer A. The column was washed isocratically for 180min and then eluted with a gradient of 0-1.0M NaCl in Buffer A over 120min. Enzymatic activity was measured by HPLC using the synthetic peptideSQNYPIV as described in Margolin et al., Biochem. Biophys. Res. Commun.,167, 554-560 (1990); the production of the pl peptide (SQNY) wasmeasured.

Active fractions were pooled, made 1.2M in ammonium sulfate, and appliedto a hexyl agarose column (2.0×18 cm) equilibrated in Buffer Acontaining 1.2M ammonium sulfate. The sample, in 125 ml (1.85 mg/ml),was loaded at a flow rate of 1 ml/min at 4° C. The column was washedwith the equilibration buffer for 240 min (1 ml/min) following sampleloading and then a reverse linear gradient of 1.2-0M ammonium sulfate inBuffer A was applied to the column over 120 min at the same flow rate.The column was then washed isocratically in Buffer A for 120 min. Activefractions were pooled, concentrated to 10 ml using an Amicon stirredcell with a YM-10 membrane, and applied at 1 ml/min to a MonoS cationexchange column (1.0×10 cm) equilibrated in Buffer A. After washingisocratically for 30 min, protease was eluted using a linear gradient of0-0.45M NaCl in Buffer A over 40 min followed by an isocratic wash withBuffer A containing 0.45M NaCl over 30 min. Chromatography was performedat 25° C.

Pooled active fractions were concentrated to 200 ul using an Amiconstirred cell and a YM-10 membrane, and the protease was then applied toa Superose 6 size exclusion column equilibrated in Buffer A containing0.1M NaCl. The column was washed isocratically in this buffer at a flowrate of 0.5 ml/min. HIV protease was eluted as a single peak.

QAE-Sepharose, and hexyl agarose were purchased from Sigma ChemicalCompany. Superose 6 and MonoS were products of Pharmacia. Buffers andreagents were obtained from Sigma.

In an analogous manner, a culture of E. coli K12 507/pHGAG was grown tomid-log phase at 32° C. then shifted to 40° C. for about 4 to 5 hours.The culture was chilled on ice and centrifuged, then the pellet wasresuspended in 8 ml lysis buffer containing 5 mg/ml lysozyme. Lysisbuffer was comprised of 50 mmol Tris-HCl (pH 7.8), 5 mmol EDTA, 1 mmolDTT, 100 mmol NaCl, 1 μg/ml E64 and 2 μg/ml aprotinin. The culture wasincubated about 30 to 60 minutes at 4° C., then briefly sonicated in aBranson® Cell Disrupter at 60% power, for three 20 second bursts withchilling between each burst. The culture was then centrifuged at 15,000g. The supernatant, which contains the unprocessed gag protein, waspartially purified by size exclusion chromatography on a Sephadex G-50column and stored at -20° C. in 50% glycerol and lysis buffer.

E. Western Blot Analysis of HIV Protease Inhibition Assay

About 10 to 20 B1 of the purified protease was preincubated with serialdilutions (100 nM-0.1 nM; five-fold serial dilutions) of the compound tobe tested at room temperature for 30 minutes. About 2 μl of the gagpolyprotein was then added and incubated at 37° C. for one hour.

Next, about 25 μl of the reaction was mixed with loading buffer andsubjected to polyacrylamide gel electrophoresis on a 12.5% Laemmli gelunder reducing conditions. The proteins were transferred onto anitrocellulose filter using standard "Western Blot" electrophoresis at500 mAmps for 35 minutes at room temperature. The filter was air-dried,then non-specific immunoreaction was blocked by washing the filter for30 minutes at 4° C. in a solution of 3% non-fat dry milk powder inphosphate buffered saline (PBS).

Monoclonal antibodies against p17 and/or p24 (available from DuPont,Wilmington, Del.) were added to about 40 ml of a 3% milk/PBS solution toa final concentration of 2 μg/ml. The nitrocellulose filter was added tothe antisera solution and slowly rocked for 2 hours at 4° C. The filterwas then removed and washed 5 times with PBS, then the maturation of thegag gene was determined by reaction with I¹²⁵ -labeled protein A or byreaction with biotinylated anti-mouse IgG antibodies (Vecta Stain®) .

To run the I¹²⁵ -labeled protein A reaction, about 10 μCi of I¹²⁵-labeled protein A was added to 100 ml of the 3% milk/PBS solution. Thefilter was rocked in this solution for 2 hours at 4° C., washed threetimes with PBS, then two times with PBS and 0.1% Tween 20. The filterwas then air-dried and exposed to x-ray film for autoradiography.

The Vecta Stain® (Vector Laboratories, Burlingome, Calif.) procedure wasperformed according to the manufacturer's recommendations. Four drops ofthe biotinylated IgG was added to 40 mls of the 3% Milk/PBS solution,then the filter was rocked in this solution for 60 minutes at 4° C. Thefilter was washed five time with PBS, then rocked 30 to 60 minutes at 4°C. in Reagent ABC. Reagent ABC was prepared by adding 4 drops of ReagentA (Avidin DH) and 4 drops of Reagent B (biotinylated peroxidase) to 20ml of 3% Milk/PBS. The filter was then washed five time with PBS, thenthe filter was developed by washing it for 5 to 10 minutes in 10 ml ofPBS containing 0.02% H₂ O₂ and 10 ml of 4-chloro-1 naphthol. When thecolor was fully developed, the filter was washed with distilled waterand air-dried.

Following the procedure, it is simple to determine the activity of theHIV-1 protease compounds of the present invention. When the protease isfully active, the gag protein, is cleaved from the precursor gag forminto mature p24 and p17 proteins, which are readily measured by theirpresence relative to the precursor protein. The addition of an HIV-1protease inhibitor to the reaction prevents the maturation of the gagprotein, an event which is easily determined using gel electrophoresisand Western Blotting.

The HIV-1 protease inhibition activity of the compounds of Formula Iwere tested using this method. The compound is diluted, then aliquots ofthe solution are added to the purified protease and the protease andcompound are "preincubated" at room temperature for 30 minutes. The gaglysate, MES buffer, DTT and NaCl were then added to the proper finalconcentrations and the reaction mixture was incubated one hour at 37° C.The reaction mixtures were then electrophoresed, "Western Blotted" anddeveloped to determine the inhibitory activity of the compounds. Theactivity of each compound was determined by the yield of p17 as measuredby LKB Ultrascan XL Laser densitometer. Using the yield of p17 in theabsence of inhibitor as 100%, the percent inhibition of the testcompounds at each dilution was determined.

The IC₅₀ results obtained in the Fluorescence Assay and Western Blot forthe compounds of the present invention are set forth below in Table 1.

                  TABLE 1                                                         ______________________________________                                        Inhibitory Activity                                                                    Fluorescence Assay IC.sub.50 *                                       Example No.                                                                            in ng/ml        Western Blot IC.sub.50 in nM                         ______________________________________                                         1       1.0             0.8                                                   2       2.1             50                                                    3       18              60                                                    4       2.9             2.7                                                   5       8.4             --                                                    6       140             700                                                   7       18              --                                                    8       3.0             1.5                                                   9       8.6             --                                                   10       59              --                                                   11       10              340                                                  12       500             >1000                                                13       8               9                                                    14       85              100                                                  15       2.7             1.6                                                  16       34              1.0                                                  17       1.2             0.5                                                  18       31              --                                                   19       2.1             --                                                   20       0.7             --                                                   21       2.5             --                                                   22       6.6             --                                                   23       0.4             --                                                   24       0.3             --                                                   25       1.0             --                                                   26       0.7             --                                                   27       5.1             --                                                   28       7.0             --                                                   29       72              --                                                   30       7.2             --                                                   31       7.1             --                                                   32       2.4             --                                                   33       1.4             --                                                   34       4.5             --                                                   35       78              --                                                   36       650             --                                                   37       21              --                                                   38       0.77            --                                                   39       3.3             --                                                   40       28              --                                                   41       8               --                                                   42       340             --                                                   43       1.3             --                                                   44       1.3             --                                                   45       0.88            --                                                   46       1.3             --                                                   47       3.4             --                                                   48       0.8             --                                                   49       0.5             --                                                   50       0.16            --                                                   51       9.2             --                                                   52       2.9             --                                                   53       0.77            --                                                   54       49              --                                                   55       0.81            --                                                   56       24.2            --                                                   57       9.7             --                                                   58       1.8             --                                                   59       7.2             --                                                   60       1.0             --                                                   61       1.3             --                                                   62       3.8             --                                                   63       0.5             --                                                   64       0.5             --                                                   65       1.2             --                                                   66       1.5             --                                                   67       5               --                                                   68       0.15            --                                                   69       0.76            --                                                   70       0.58            --                                                   ______________________________________                                         *IC.sub.50 values for Examples 2-70 are standardized relative to Ex 1.   

We claim:
 1. A compound having the formula ##STR12## where R¹ is aryl orC₅ -C₇ cycloalkyl;R^(b) is hydrogen or an amino protecting group; Y isphenyl, substituted phenyl, naphthyl, substituted naphthyl, quinolinyl,furyl, phienyl, benzofuryl, or benzothienyl; and R³ is a group havingthe structure; ##STR13## I is 3, 4, or 5; p is 4 or 5; R⁴ at eachoccurrence is independently hydrogen, C₁ -C₆ alkyl, or hydroxy (C₁ -C₄)alkanediyl; R⁵ and R⁶ are independently selected from hydrogen, hydroxy,C₁ -C₆, alkyl, C₁ -C₆ alkoxy, amino, C₁ -C₄ alkylamino, hydroxy (C₁-C₄)-alkanediyl, carboxy, (C₁ -C₄ alkoxy)carbonyl, aminocarbonyl, C₁ -C₄alkylaminocarbonyl or aryl or a pharmaceutically acceptable salt orsolvate thereof.
 2. A process for preparing a compound of claim 1comprising:a) reacting a compound having the Formula: ##STR14## where R³and Y are as defined above with a C₁ -C₄ alkyl lithium or lithium di(C₁-C₄ alkyl) amide base optionally in the presence of a tetramethyl(C₁ -C₄alkylene)diamine catalyst in an aprotic solvent to afford thecorresponding anion; and b) reacting the anion from (a) with an amidehaving the Formula ##STR15## where R^(b) and R¹ are as defined above inan aprotic solvent.